Discovery and Characterization of Unusual O-Linked Glycosylation of IgG4 Antibody Using LC-MS.

ABSTRACT:

Background

Consensus is that immunoglobulin IgG4 contains only N-linked glycosylation. The analysis of several batches of commercial biopharmaceutical product Dupixent using top-down intact mass spectrometry revealed that this IgG4 features a small amount of O-linked glycosylation in the Fab region. This is the first report of an O-linked glycosylation in an IgG4 antibody.

Methods

Monoclonal antibody solutions were subjected to cation exchange (CEX) and reverse phase (RP) chromatography and/or additional preconcentration/fractionation methods to prepare samples for subsequent analysis. Advanced MS analysis and fragmentation techniques (HCD, ETD, and EThcD) were employed to localize the O-linked glycosylation as well as elucidate the structure of the glycan(s).

Results

SUBMITTER: Janecki DJ 

PROVIDER: S-EPMC11635057 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature