title: Highly sensitive capillary electrophoresis analysis of N-linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser-induced fluorescence detection.
abstract: We describe a highly sensitive CE with laser-induced fluorescence (LIF) detection for the analysis of N-linked oligosaccharides in glycoproteins using rhodamine 110 as a fluorescence derivatization reagent. One CE separation is performed using a fused-silica capillary and neutral pH buffer conditions and allows for the separation of sialo-oligosaccharides according to the number of sialic acids. An alternate separation is performed using the same capillary and acidic pH buffer conditions, enabling the separation of asialo-oligosaccharides according to their sizes. The derivatization and separation conditions for the analysis of sialo- and asialo-oligosaccharides were optimized. Furthermore, we applied the proposed method for the analyses of N-linked sialo- and asialo-oligosaccharides in glycoproteins (ribonuclease B, fetuin, and recombinant human erythropoietin).
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, Johnan, Fukuoka, Japan.
authors.forename: Soichiro
authors.initials: S
authors.lastname: Ijiri
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Todoroki
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Yoshida
authors.forename: Takashi
authors.initials: T
authors.lastname: Yoshitake
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Nohta
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Yamaguchi
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
chemical_list: D005456:Fluorescent Dyes; D006023:Glycoproteins; D009844:Oligosaccharides; D012235:Rhodamines; C049025:rhodamine 110