Peptide Mapping

Understand primary protein structure and assess post translational modifications

Graphical description of PTMs

Peptide Mapping of Biosimilars

Peptide Mapping Service

  1. Reduction and alkylation of the protein 
    • Reduction of S-S bridges and blocking of the generated free thiol opens the protein structure and allows proteolytic digestion to be more efficient. In the case of multi-chain proteins which have disulfide bridges linking the chains (e.g. monoclonal antibodies) this step will also break the bridges between the chains.
  2. Digestion of the protein using specific enzymes
    • This step breaks the protein down into peptides. Enzymes  are chosen based on the theoretical sequence so this informed choice of enzymes(s) results in the best peptide mapping analysis.
  1. Analysis of the digested peptides using on-line RP-HPLC with UV and LC/ES-MS.
    • LC separates the peptides based on polarity (reverse phase-LC).
    • The Mass Spec generates mass information for each peptide as it elutes from the LC column. Fragment ion information is also generated, allowing the primary sequence to be determined which serves to confirm peptide identity. The mass spectrometers at BioPharmaSpec are also capable of generating fragmentation information from the peptides as they enter the MS source from the LC column. This is either selective (MS/MS) or non-selective (MSe), depending on the requirements of the study and can be considered as a form of tandem mass spectrometry.

Multiple Applications of Peptide Mapping

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