Comparison of N-Glycopeptide to Released N-Glycan Abundances and the Influence of Glycopeptide Mass and Charge States on N-Linked Glycosylation of IgG Antibodies

• DOI: 10.1021/acs.jproteome.3c00904
• Publication Date: 2024-03-07T12:17:29Z
• Authors: Concepcion A. Rem...; Meghan C. Burke; Tytus D. Mak; Sergey L. Sheetlin; Yuri A. Mirokhin; Brian T. Cooper; Zachary C. Goecker; Mark S. Lowenthal; Xiaoyu Yang; Guanghui Wang; Dmitrii V. Tchekh...; Stephen E. Stein
• Keywords: Fragmentation

We report the comparison of mass-spectral-based abundances tryptic glycopeptides to fluorescence released labeled glycans and effects mass charge state in-source fragmentation on glycopeptide abundances. The primary glycoforms derived from Rituximab, NISTmAb, Evolocumab, Infliximab were high-mannose biantennary complex galactosylated fucosylated N-glycans. Except for ions loss HexNAc or HexNAc-Hex sugars are prominent other therapeutic IgGs. After excluding results, a linear correlation was observed between fluorescently N-glycan over dynamic range 500. Different states human IgG-derived containing wider variety abundant attached also investigated examine ion These revealed dependence abundance glycan with higher favoring higher-mass glycans. Findings indicate that spectrometry-based bottom-up approach can provide results as accurate those release studies while revealing origin each glycan. site-specific relative conveniently displayed compared using previously described distribution spectra "GADS" representations. Mass spectrometry data available MAssIVE repository (MSV000093562).