Before approval by regulatory agencies (FDA, EMA and others), biotherapeutic proteins require an extensive characterization. One of the requirements is the primary structure confirmation, which can be performed by liquid chromatography coupled to mass spectrometry (LC-UV-MS). Due to the costs of a MS analysis and the lack of equipment for routine quality control, methods are often developed on LC-UV-MS systems and QC samples are analyzed by LC-UV only. However, interfering peaks can be observed in QC, generated by trypsin non-specific activity or autoproteolysis, and lead to unwanted investigations.
This poster showcases the advantages of the newly developed trypsin compared to commercially available trypsins. We are also focused on the benefits of the replacement of the currently used trypsin with the new trypsin in Trypsin/Lys-C mix, which is commonly used to digest proteolytically resistant sites. Finally, we have confirmed the use of the new trypsin with AccuMAP™ Low pH Protein Digestion kit to characterize deamidation, a critical quality attribute tracked during multiple attribute monitoring experiments.
New trypsin significantly improves characterization of biotherapeutics:
- It suppresses non-specific, chymotryptic-like cleavages commonly induced by commercially available trypsins;
- Combination of new trypsin with Lys-C provides an efficient tool for digestion of proteolytically resistant cleavage sites;
- AccuMAP™ kit supplemented with new trypsin allows for effective and accurate characterization of deamidation.