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Analysis of carbohydrates in glycoproteins by high-performance liquid chromatography and high-performance capillary electrophoresis

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Abstract

We describe two methods for the analysis of oligosaccharide chains in glycoproteins by high-performance liquid chromatography (HPLC) and high-performance capillary electrophoresis (HPCE).O-andN-glycosidically linked oligosaccharides released from glycoproteins can be identified as their borohydride-reduced forms by anion-exchange HPLC with pulsed amperometric detection.N-Glycosidically linked oligosaccharides can also be analyzed as 2-aminopyridine derivatives by HPCE in direct zone electrophoresis mode in an acidic phosphate buffer and zone electrophoresis mode as borate complexes in an alkaline buffer.

We also present a convenient procedure for the analysis of the constituent monosaccharides of these oligosaccharides chains by HPLC based on reversed-phase partition mode as 1-phenyl-3-methyl-5-pyrazolone derivatives.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Kinki University, Higashi-Osaka, Japan

    Kazuaki Kakehi & Susumu Honda

Authors
  1. Kazuaki Kakehi
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  2. Susumu Honda
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Additional information

This article first appeared: in Hounsell, E. F., ed. (1993),Methods in Molecular Biology, Vol. 14: Glycoprotein Analysis in Biomedicine, Humana Press Inc., Totowa, NJ.

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Kakehi, K., Honda, S. Analysis of carbohydrates in glycoproteins by high-performance liquid chromatography and high-performance capillary electrophoresis. Appl Biochem Biotechnol 43, 55–71 (1993). https://doi.org/10.1007/BF02916431

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  • DOI: https://doi.org/10.1007/BF02916431

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