N-Glycan Analysis by Ultra-Performance Liquid Chromatography and Capillary Gel Electrophoresis with Fluorescent Labeling

Curr Protoc Protein Sci. 2019 Sep;97(1):e95. doi: 10.1002/cpps.95.


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.

Keywords: N-glycan; capillary gel electrophoresis; fluorescent labeling; liquid chromatography; posttranslational modification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods
  • Electrophoresis, Capillary / methods
  • Fluorescent Dyes / chemistry
  • Glycoproteins / chemistry
  • Plasma / chemistry
  • Polysaccharides / analysis*
  • Protein Denaturation
  • Reproducibility of Results
  • Spectrometry, Fluorescence / methods
  • Staining and Labeling / methods


  • Fluorescent Dyes
  • Glycoproteins
  • Polysaccharides