Rapid analysis of oligosaccharides derived from glycoproteins by microchip electrophoresis

Fuquan Dang, Kazuaki Kakehi, Kazuki Nakajima, Yasuo Shinohara, Mitsuru Ishikawa, Noritada Kaji, Manabu Tokeshi, Yoshinobu Baba

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24 Citations (Scopus)

Abstract

A novel method for fast profiling of complex oligosaccharides released from glycoproteins based on microchip electrophoresis (μ-CE) is presented here. The characterization of separation conditions, i.e., the composition, concentration and pH of running buffer as well as the applied voltage, has been performed using maltose (G2), cellobiose (G′2), maltriose (G3) and panose (G′3) as oligosaccharide isomer models. In μ-CE, much better separation of oligosaccharide isomers and oligosaccharide ladder was obtained in phosphate buffer than in borate buffer over a wide pH range. Under optimal conditions, high-performance separation of the N-linked complex oligosaccharides released from ribonuclease B, fetuin, α1-acid glycoprotein (AGP) and IgG was achieved using polymethylmethacrylate (PMMA) microchips with an effective separation channel of 30 mm. These results represent the first reported analysis of the N-linked oligosaccharides derived from glycoproteins by μ-CE, indicating that the present μ-CE-based method is a promising alternative for characterization of the N-linked oligosaccharides in glycoproteins.

Original languageEnglish
Pages (from-to)138-143
Number of pages6
JournalJournal of Chromatography A
Volume1109
Issue number2
DOIs
Publication statusPublished - 24-03-2006

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All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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