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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:
In our Mass Spectrometric Peptide Mapping services, we cleave the protein/antibody into smaller peptides using a specific protease. The peptides are analyzed by mass spec and then the observed peptides correlated to the protein/antibody amino acid sequence for peptide identification.
The identified peptides in the peptide map thus confirms the specific amino acid sequences covered by the peptide map, as well as the identity of the protein/antibody. 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%.
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.
Peptide mapping analysis of mAb
The peptide mapping using high-flow LC-MS/MS is performed on a Agilent 1200 system running at 0.2 ml/min equipped with 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 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.
Antibody sequence coverage map
Protein/Antibody peptide mapping analysis by mass spectrometry requires a pure protein/antibody in sufficient amounts to obtain good data. It is thus essential to prepare samples in a clean laboratory to avoid contamination with human keratin.
We suggest that you provide samples with a chromatographic purity of minimum 80%. Let us know if you have glycerol, detergents, or high salt content in your samples.
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.
