Glycosylation is well-recognized as a critical quality attribute (CQA) or product quality attribute (PQA) of biotherapeutics, and is routinely monitored to ensure desired product quality, safety, and efficacy.  Additionally, as one of the most prominent post-translational modifications, glycosylation plays a key role in disease manifestation, and changes in glycosylation may serve as specific and sensitive biomarkers. However, glycans are highly heterogeneous, which has considerably hampered the progress of glycomics. This webcast will describe complete N-glycan quantitation workflows using the Agilent AdvanceBio InstantPC label for high-throughput and high-resolution glycan profiling using hydrophilic interaction liquid chromatography (HILIC) with detection by fluorescence (FLD) or mass spectrometry (MS).

Topics covered:

• Rapid and reproducible N-glycan preparation from glycoproteins of varied sources (1 hour for InstantPC, 2 hours for 2-AB)
• Relative quantitation of released N-glycans in a reliable and high-resolution way (FLD and MS detection)
• Characterization of InstantPC N-glycans by exoglycosidase sequential digestion
• Detailed workflow for construction and use of InstantPC glycan glucose unit (GU) database
• Innovative 2D liquid chromatography for fraction and separation of complex InstantPC N-glycans