TY - JOUR
T1 - Rapid analysis of oligosaccharides derived from glycoproteins by microchip electrophoresis
AU - Dang, Fuquan
AU - Kakehi, Kazuaki
AU - Nakajima, Kazuki
AU - Shinohara, Yasuo
AU - Ishikawa, Mitsuru
AU - Kaji, Noritada
AU - Tokeshi, Manabu
AU - Baba, Yoshinobu
N1 - Funding Information:
The present work is supported in part by the CREST program of the Japan Science and Technology Corporation (JST); a grant from the New Energy and Industrial Technology Development Organization (NEDO) of the Ministry of Economy, Trade and Industry, Japan; a Grant-in-Aid for Scientific Research from the Ministry of Health and Welfare, Japan; and a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Technology, Japan.
PY - 2006/3/24
Y1 - 2006/3/24
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/33644822835
UR - https://www.scopus.com/pages/publications/33644822835#tab=citedBy
U2 - 10.1016/j.chroma.2005.11.133
DO - 10.1016/j.chroma.2005.11.133
M3 - Article
C2 - 16376899
AN - SCOPUS:33644822835
SN - 0021-9673
VL - 1109
SP - 138
EP - 143
JO - Journal of Chromatography A
JF - Journal of Chromatography A
IS - 2
ER -