Mapping IgG Subunit Glycoforms Using HILIC and a Wide-Pore Amide Stationary Phase

Monoclonal Antibodies (mAbs) are often characterised by proteolyzing using IdeS and identifying using reversed phased (RP) chromatography, however modification of residues in the protein can dramatically effect RP retention times. This application note suggests the use of hydrophilic interaction chromatography (HILIC) with an amide bonded stationary phase to separate IgG proteins with N-linked glycosylation. This article highlights the capabilities of the glycoprotein BEH amide, 300Å, 1.7 μm to characterise mAb glycosylation, including elucidation of domain-specific glycan information.