Wang HJ, O'Grady J, Horn D, Hao Z, Meyer K, Josephs J.
ASMS 2014 Poster
Purpose: To develop a workflow for fast and comprehensive characterization of peptide sequence, identification and relative quantitation of post-translational modifications (PTMs) of monoclonal antibody (mAb). Methods: Native and oxidatively stressed IgG mAb were enzymatically digested by Flash DigestTM kit with trypsin. Peptide samples were analyzed by online LC-MS on a Thermo Scientific™ Q Exactive™ mass spectrometer. Peptide sequence mapping, identification and quantification of PTMs were performed by Thermo Scientific™ PepFinderTM software (version 1.0). Results: Comprehensive and simultaneous sequence and PTM analysis with a particular focus on oxidation of IgG mAb was realized by combining rapid digestion, high resolution, accurate mass (HRAM) data and PepFinder software. This workflow greatly shortens the sample preparation and data analysis time while providing great sensitivity to detect low level PTMs.
Thermo Fisher Scientific, Perfinity Biosciences Inc.
United States
