Peptide Mapping

Peptide mapping is a powerful tool for understanding how a protein may be modified to suit your purpose. It provides a whole suite of information to help promote the drug development process. Creative Biolabs now provides peptide mapping service that allows you to meet the structural characterization requirements of the specified guidelines, understand the primary structure of your antibody candidate, and make key drug development decisions based on this knowledge.

Antibody Peptide Mapping

Peptide mapping is a key tool for pharmaceutical research and drug development to bioproduction and clinical testing. Antibody peptide mapping involves selective cleavage of the protein to generate smaller peptides of characteristic, reproducible sizes. Then, the peptides are separated on a gel to determine their masses or they undergo other types of mass characterization techniques like mass spectrometry (MS).

Peptide mapping is commonly undertaken after cleavage of the antibody into proteolytic peptides. When using one digestion enzyme such as trypsin, peptide mapping is typically carried out for protein identification. The analysis is performed using matrix-assisted laser desorption ionization (MALDI)-MS/MS or liquid chromatography-electrospray ionization-tandem MS (LC-ESI-MS/MS), for example during protein identification after electrophoresis, such as 1D or 2D PAGE. When using several enzymes together, peptide mapping is applied for the confirmation of a complete amino acid sequence, for example when confirming the amino acid sequence of a biosimilar and comparing it with the originator molecule. In order to obtain the full sequence information of the sample, Creative Biolabs uses up to 6 kinds of proteases for protein digestion (Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C, and Lys-N). After protein digestion, sequences of peptide are mapped for a full-length sequence of biopharmaceuticals.

Depending on the experimental setup, peptide mapping can also be used to determine the N- and C-terminus of a protein. This is extremely crucial information for our clients in the case of monoclonal antibodies (mAbs), where the truncation level of the C-terminal Lysine can be monitored. In combination with additional sample preparation, peptide mapping can also be used to analysis:

• The degree of deamidation or oxidation for each affected amino acid
• Glycosylation sites and structure
• N-glycosylation sites
• Disulfide linkages
• N-terminal pyroglutamic acid
• C-terminal lysine clipping.

Peptide Mapping Analysis Workflow

• Proper enzymes are selected for enzyme digestion and identification of target proteins.
• Peptide analysis through LC-MS/MS or UV LC-MS/MS.
• The total sequence of peptide coverage/peptide is mapped and compared with the theoretical sequence.
• Delivery of report.

Advantage of Our Peptide Mapping Services

• Combination of specific and nonspecific proteases, achieving coverage of 100% of antibody.
• Confirmation of each peptide is based on the MS/MS fingerprint of the sample and is more accurate than MS analysis.
• The identification is based on LC-ESI-MS/MS data, which allows the sequencing of peptides with high confidence.
• Overlapping peptide sequences confirm the order of amino acids in the sequence.

Reports

• Experiment procedures
• Parameters of LC and MS
• MS raw data files
• Peptide mapping results
• Bioinformatics analysis

Creative Biolabs offers comprehensive peptide mapping services at the core of our expertise. We work internationally with a wide range of life science businesses. As such, we are the perfect partner to provide your business with antibody fingerprinting solution. Contact us now to find out how our peptide mapping analysis can be applied to your drug development project.

For Research Use Only.