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Glycan Standards

BioFiles 2007, 2.1, 18.

BioFiles 2007, 2.1, 18.

The analysis of the glycan portion of glycoproteins is necessary for monitoring changes in post-translational modification that occur in disease states, as well as evaluating the consistency of glycoprotein production. In general, the method is to isolate glycan pools from gel electrophoresis bands of a glycoprotein using in-gel digestion techniques.1 The isolated glycan pools are subsequently separated by a chromatographic method, usually high performance liquid chromatography (HPLC), high-pH anionexchange chromatography (HPAEC), hydrophilic interaction liquid chromatography (HILC), or high-pH anion-exchange chromatography (HPAE).2 Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry analysis of the glycan constituents is used to identify the individual glycans after separation.3,4,5

Glycan standards function as markers during the separation and purification of glycans isolated from glycoproteins. These compounds are used as internal reference compounds for the peak assignment of glycan constituents separated by chromatography. Additionally, they may be used for calibration in MALDI-TOF mass spectrometry analysis. The following are the most common N-linked and O-linked glycans for research applications.

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Materials

     

References

  1. Rudd, P. and Dwek, R., Rapid, sensitive sequencing of oligosaccharides from glycoproteins, Curr. Opin. Biotechnol., 8, 488 (1997).
  2. Charlwood, J. et al., Characterization of the glycosylation profiles of Alzheimer’s β-secretase protein Asp-2 expressed in a variety of cell lines, J. Biol. Chem., 276, 16739 (2001).
  3. Kremmer, T., et al., Liquid chromatographic and mass spectrometric analysis of human serum acid α-1-glycoprotein, Biomed. Chromatogr., 18, 323 (2004).
  4. Yu, X., et al., Identification of N-linked glycosylation sites in human testis angiotensin-converting enzyme and expression of an active deglycosylated form, J. Biol. Chem., 272, 3511 (1997).
  5. Leibiger, H. et al., Structural characterization of the oligosaccharides of a human monoclonal anti-lipopolysaccharide immunoglobulin M, Glycobiology, 8, 497 (1998).

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