Rapid analysis of oligosaccharides derived from glycoproteins by microchip electrophoresis.
A novel method for fast profiling of complex oligosaccharides released from glycoproteins based on microchip electrophoresis (mu-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 ( G2'), maltriose (G3) and panose (G3') as oligosaccharide isomer models. In mu-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, alpha1-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 mu-CE, indicating that the present mu-CE-based method is a promising alternative for characterization of the N-linked oligosaccharides in glycoproteins.
Dang
Fuquan
F
Health Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho 2217-14, Takamatsu 761-0395, Japan.
Kakehi
Kazuaki
K
Nakajima
Kazuki
K
Shinohara
Yasuo
Y
Ishikawa
Mitsuru
M
Kaji
Noritada
N
Tokeshi
Manabu
M
Baba
Yoshinobu
Y
D002240:Carbohydrate Sequence; D046209:Electrophoresis, Microchip / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis*
D006023:Glycoproteins; D009844:Oligosaccharides