The analysis of complex N- and 0-linked glycans composed of frequently similar and repeating sugar moieties requires a highly discriminating chromatographic method. Hydrophilic liquid interaction chromatography (HILIC), using UPLC, HPLC, or UHPLC columns containing Waters amide bonded, Ethylene Bridged Hybrid Technology (BEH) particles separates compounds based on differing hydrophilicity.
Released N-Glycan Columns
Waters ACQUITY UPLC and XBridge Glycan BEH Amide 130Å Columns are best suited for the analysis of released, N-labeled glycans using pre-column labeling with 2-AB, 2-AA, or Waters innovative and enabling RapiFluor-MS reagent. Each batch our Glycan BEH Amide 130Å chemistry is specifically QC tested with a complex mixture of 2-AB labeled glycans from a mAb digest to help ensure consistent column performance in validated methods.
The image to the right shows the N-linked glycans from pooled human IgG contain high mannose as well as neutral, and sialylated complex structures. The chromatogram shows 35 minutes of a one hour analysis. This challenging sample includes high mannose, bisecting GlcNAc and sialylated species.
Shown to the right is the performance of HILIC glycan columns varying in particle size at the same linear velocity.