Peptide mapping services
- Confirm the amino acid sequence of proteins and antibodies
- Quantify PTMs
- Check for amino acid substitutions and identify degradation products
We offer customized solutions, contact us to discuss your project.
Send us a description of your project and we shall be happy to present you with a solution.
We encourage you to call us or schedule a meeting to discuss your project in detail.
Set up meetingProtein characterization for optimization of manufacturing processes.
Explore customer caseCombining mass spec analyses for mAb characterization and batch-to-batch comparison
Explore customer caseWe would like to help you as much as possible with your project and therefore provide several kinds of customer support:
We cleave the protein/antibody into smaller peptides using a specific protease in our Mass Spectrometric Peptide Mapping services. The peptides are analyzed by mass spec, and then the observed peptides are correlated to the protein/antibody amino acid sequence for peptide identification.
Peptide mapping using a single protease typically gives a protein sequence coverage of 30-90%. By using two or more proteases, this protein coverage increases to approx. 95-100%. The identified peptides in the peptide map thus confirm the specific amino acid sequences covered by the peptide map and the protein/antibody identity.
Detailed characterization of a protein/antibody requires careful investigation of the protein/antibody sequence to select the best proteases by mass spec. We use UV LC-MS/MS to observe the peptides.
The peptide mapping using high-flow LC-MS/MS is performed on an Agilent 1200 system running at 0.2 ml/min equipped with a C18 column connected to a high-resolution Q-TOF instrument (Bruker Impact HD). The instrument runs either with a short or long gradient, and the Q-TOF is set to data-dependent analysis mode (DDA), switching between MS and MS/MS mode.
The analysis provides both a UV- and a Base peak chromatogram, and all MS/MS data is collected for peptides with m/z from 200 to 1600 Da.
Protein/Antibody peptide mapping analysis by mass spectrometry requires sufficient amounts of pure protein/antibody to obtain good data. It is thus essential to prepare samples in a clean laboratory to avoid contamination with human keratin.
We suggest you provide samples with a minimum of 80% chromatographic purity. Let us know if your samples have glycerol, detergents, or high salt content.
Provide samples in solid form or in solution. Reach out if you have gel bands you would like to analyze.
Minimum sample amount: ~ 100 microgram protein and ~ 0.5 milligram antibody.