Large-Scale Identification and Quantification of Covalent Modifications in Therapeutic Proteins

摘要: Covalent modifications on therapeutic proteins are traditionally monitored by chromatographic techniques, which quantify limited number of protein at a time. In this report, computer algorithms for automated analyses liquid chromatography/tandem mass spectrometry (LC/MS/MS) data large-scale identification and quantification known unknown described. Peptide is achieved comparing the experimental fragmentation spectrum to predicted each native or modified peptide. Peak areas related peptide ions under their selected-ion chromatograms (SIC) used relative peptides. A matched window function generate SIC more reliable quantification. an LC/MS/MS analysis tryptic digestion IgG2 monoclonal antibody, 1712 were identified with false-discovery rate approximately 0.4%, 227 quantified. The accuracy spectrometry-based evaluated abundance different glycoforms determined that fluorescence-based chromatography method. This method may potentially replace many methods assessing quality attributes proteins.