Fine characterization of therapeutic antibody Trastuzumab by UHPLC-ESI-QTOF-MSE peptide mapping using reversed-phase and HILIC separation modes

Monoclonal antibodies are among the most promising molecules for the pharmaceutical industry, but their characterization remains very challenging due to their complexity. Although mass spectrometry is now a well established technique for development work,some people are still reluctant to use it for batch release, as it is believed to be expensive and time consuming. In the present work, we present peptide mapping methodologies that can be easily used to characterize monoclonal antibodies. Tryptic peptides are usually separated by reversed-phase chromatography before UV and/or MS detection. We show that HILIC mode (Hydrophilic Interaction Liquid Chromatography) can be an interesting alternative, especially for an effi cient separation of glycopeptides.