The presence and abundance of glycans post translationally conjugated to the monoclonal antibody influence a number of characteristics such as molecule structure, effector function and stability, and as such are a serious challenge to developers of antibody therapeutics.
Released N-Glycan Analysis by LC/MS
BioOutsource offers a released N-Glycan assay for the in-depth profiling of IgG glycosylation. Glycans are enzymatically removed from the antibody and then labelled before a high resolution LC separation is performed as demonstrated in Figure 1. Fluorescence detection coupled with online ESI mass spectrometry enables confident assignment and accurate quantification of the different glycan structures. These can then be grouped according to their structural classification (e.g. complex, oligomannose) and monosaccharide composition, such as fucose, galactose and sialic acid profiles.
The chromatogram above shows the main glycoforms identified, relative to the most abundant GOF peak set to 100%.
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