2P-48 PDF
Peptide mapping of biotherapeutic proteins by automated data processing on high resolution accurate mass spectrometry with SWATH® acquisition
Analytical Technology and the related informatics have advanced rapidly in response to a highly dynamic biotechnology market. In some of the most responsive segments, the ability to determine the identity of peptides and then confirm that identity is a key characteristic in developing business efficiency.
Biotherapeutic proteins carry more complexity than the traditional small molecule drugs. The heterogeneity assessments are vital to ensure product quality – drug efficacy and patient safety. Mass spectrometry techniques are a powerful tool to answer the critical questions: full amino acid sequence, post-translational modification (PTM) ratios, as well as possible process and storage derived modifications.
Here we utilize for peptide mapping for the first time SWATH® acquisition, a data independent acquisition (DIA) technique, unique to SCIEX Q-TOF instrumentation. The SWATH technique overcomes one of the issues associated with the default data acquisition methods: peptides (precursor ions) being irreproducible picked for MS/MS analysis. With SWATH acquisition all peptides are selected for MS/MS analysis – without instrument (data-dependent) or prior user (targeted) precursor ion selection. The data acquisition in SWATH acquisition mode enabled all information to be collected, including the lower abundant features that could easily be missed by usual data-dependent acquisition modes.