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Released N-glycan profiling

Glycosylation is one of the most frequent protein post-translational modifications, which may affect the stability, immunogenicity, pharmacokinetics and biological activity of the protein. Therefore, comprehensive characterization and monitoring of glycans is an essential task in development of biologics. Characterization of released glycans provides more comprehensive information about the glycan profile in comparison to analysis of the intact glycoprotein.

In the released N-glycan profiling, the protein is first deglycosylated to release N-glycans, which is followed by labelling, purification, and detection of the fluorophore labelled N-glycans. N-glycans are separated using chromatography and detected based on the sensitive fluorescence signal. A glycan library is used to identify the glycan structures based on their retention time and identification is confirmed with MS.