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Updated: Jun 17, 2026

Chemo-enzymatic Synthesis of N-glycans for Array Development and HIV Antibody Profiling
Published on: February 5, 2018
Separation of 2-aminobenzamide labeled glycans using hydrophilic interaction chromatography columns packed with 1.7
Joomi Ahn1, Jonathan Bones, Ying Qing Yu
1Biopharmaceutical Sciences, Waters Corporation, 34 Maple Street, Milford, MA 01757, USA.
Abstract:
Separation by hydrophilic interaction chromatography (HILIC) with fluorescence detection utilizing a sub-2 microm glycan column for the separation of 2-aminobenzamide (2-AB) labeled N-linked glycans is described. The HILIC column packed with a 1.7 microm amide sorbent improves the peak capacity compared to a 3.0 microm HILIC column by a similar degree as observed in reversed-phase ultra-performance liquid chromatography (RP-UPLC). The results indicated that the optimal peak capacity was achieved at flow rate 0.2-0.5 mL/min. HILIC method transfer guidelines were shown to further enhance the resolution of glycans by changing initial gradient conditions, flow rate, column temperature, and different column lengths. Additionally, excellent resolution can be achieved in the separation of 2-AB labeled glycans released from fetuin, RNase B, and human IgG with a rapid analysis time.
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