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Updated: Jan 19, 2026

Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Published on: January 18, 2020
N-Glycan Analysis by Ultra-Performance Liquid Chromatography and Capillary Gel Electrophoresis with Fluorescent
Maja Hanić1, Gordan Lauc1,2, Irena Trbojević-Akmačić1
1Genos Glycoscience Research Laboratory, Zagreb, Croatia.
Abstract:
Glycans are a class of macromolecules essential for all forms of life. They embellish various proteins and other macromolecules in organisms and are responsible for their proper functioning. Because their complex structure is determined by genetic and environmental factors, analysis of such molecules is rather demanding. Liquid chromatography (high-performance and ultra-performance, HPLC and UPLC, respectively) analysis has been used for the purpose of glycoprofiling for years and it is a well-established method regarding its robustness, reproducibility, and high throughput. Another orthogonal method that is now used in glycoprofiling is capillary gel electrophoresis (CGE) because it offers powerful separation and distinct sensitivity. The purpose of the following protocols is to present all steps required for release and fluorescent labeling of total N-glycans from blood plasma/serum or isolated glycoprotein (for example, IgG) and their subsequent UPLC or CGE analysis. © 2019 by John Wiley & Sons, Inc.
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Capillary Electrophoresis (CE)
Capillary electrophoresis (CE) is a separation technique that separates molecules in an electric field according to size and charge. CE is performed in a small glass tube called a capillary that is filled with an electrolyte solution. Analytes are separated due to differences in electrophoretic mobility, which varies with charge, solvent viscosity, and size. Traditional electrophoresis in gels is limited in the amount of voltage...