Waters and Ireland’s National Institute for Bioprocessing Research and Training (NIBRT) have made available the world’s first database for glycan analysis by UltraPerformance Liquid Chromatography (UPLC).

The new GlycoBase 3+ Database developed by Professor Pauline Rudd’s research team at NIBRT,  is the first-of-its-kind repository of chromatographic retention positions expressed in glucose units for sets of glycan structures associated with a range of biotherapeutics.

The database is intended to give biopharmaceutical manufacturers a timely and powerful tool for confirming the structure of various glycosylated proteins. It has accurate information about glycosylation during the various stages of the manufacturing process and  biopharmaceutical manufacturers can gain a greater degree of control over their production process in line with regulatory guidelines aimed at guaranteeing safe and efficacious bio-therapeutics.

Recent advances in chromatographic approaches are leading to better resolution, sensitivity, and speed during the qualitative and quantitative analysis of protein glycosylation. As a proven approach for analysing biomolecules and the glycan structures attached to them, UPLC is a potent tool for determining the relative proportions of each glycan structure in a pool.

The WatersUPLC Glycan Analysis Solution features an ACQUITY UPLC BEH Glycan Separation Technology column and an Acquity UPLC System with fluorescence detection (FLR) to separate the released glycans of biopharmaceuticals as their 2-aminobenzamide (2-AB) derivatives. The UPLC Glycan Separation Solution provides robust, high-resolving, reproducible, and rapid methods that outperform analyses by HPLC.

The combination of NIBRT’s database together with the Waters UPLC Glycan Solution can readily assign potential glycan structures - complex, neutral, high-mannose, and sialylated - to each UPLC peak by matching to a dextran hydrolysate ladder (GU values). Exo-glycosidase array digestion products can also be assigned using the same bioinformatics platform, allowing the monosaccharide sequences and linkages of the oligosaccharides to be confirmed. The technology can be tailor-made to confirm structures present in samples analysed routinely or to identify unknown or unexpected glycans.