Benefits of Rapid HDX-MS Epitope Mapping
- Identify linear, conformational and structural epitopes – understand complex binding
- Resolution – higher than other methods
- Speed – answers in weeks, not months
- Cost effective – make it part of your routine
Why MS Epitope Mapping?
Epitope mapping is required for FDA and EMEA regulatory filings. It is also critical for IP protection and highly insightful for guided antibody engineering and development.
Mass spectrometry (MS) based antibody-epitope mapping service is the high throughput, high-resolution method of choice for rapid binding site characterization in therapeutic and diagnostic antibody development.
Reproducibility Impacts.
Applications of HDX-MS
While there are many possible applications of HDX-MS, Rapid Novor is highly focused on:
Antibody-Antigen epitope mapping studies
Augmenting epitope binning studies (e.g. via SPR)
Biosimilar characterization and comparability
Guided antibody discovery
Batch-to-batch reproducibility for antibody production
Antibody engineering and optimization
Who uses Epitope Mapping.
Researchers, Antibody Users
Researchers use tool antibodies every day, across many applications. Sequence confirmation is the first step in troubleshooting, or daily quality checks to prevent issues.
Antibody Venders, Producers
Easily verify each new lot. Offer premium Verified by MATCHmAb™ products, or an add-on service.
Antibody Engineering
Cell line instability could introduce mutations to humanized antibodies. Verify the CDRs have not changed or confirm the full expressed sequence matches the engineered one.
CRO, Custom Developers
Checkpoints to increase confidence. Avoid troubleshooting re-runs with clients.
How Epitope Mapping Works.
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 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.
Workflow:
- 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
| MATCHmAb™ | |
|---|---|
| Workflow | LC-MS/MS, single or dual digest |
| Bioinformatics | Real-time analysis, purpose built algorithms |
| Speed | Fast: 1-4 days |
| Throughput | High: > 100 per week |
| Coverage | Higher, and amino acid level |
| Traditional Peptide Mapping | |
|---|---|
| Workflow | LC-MS only, single enzyme only |
| Bioinformatics | Post-processing, off the shelf tools |
| Speed | Slow: weeks |
| Throughput | Low |
| Coverage | Lower, and only peptide level |
Comparison With Other Methods.
| Rapid Novor HDX | Peptide Arrays and Peptide Scanning | Crystal Structure |
|---|---|---|
| 6 Weeks | 3-10 weeks (peptide panel synthesis takes time) | Months |
| Conformational, linear, structural | Linear mostly | Conformational, linear, structural |
| 300μg of sample | 50-100μg of sample | |
|
Starting at |
$1000 to $15,000 | $$$$ |
Getting Started is Easy.
Here’s what we need:
- 20 μM of each protein solution – e.g. for antibodies 300μg of the protein in solution. Any species any isotype
- Protein sequences (we can perform sequencing if required)
- Tell us what you hope to achieve
- We’ll provide a timely quote
Deliverables
- Report of regions showing relative deuterium uptake on the sequence and on a 3D structure
- Indication of the epitope region
Talk to Our Scientists.
We Have Sequenced 6000+ 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.
Antibody Sequencing, Discovery and Proteomics Services.
Antibody Sequencing & Discovery Services.
REmAb®
Monoclonal antibody sequencing from small protein samples, no need for hybridoma or DNA information. Full sequence in record time.
Explore REmAb
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
We Have Sequenced 5000+ 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