High-resolution capillary gel electrophoresis of reducing oligosaccharides labeled with 1-aminopyrene-3,6,8-trisulfonate

被引:147
作者
Guttman, A
Chen, FTA
Evangelista, RA
Cooke, N
机构
[1] Beckman Instruments, Inc., Fullerton, CA 92634
关键词
D O I
10.1006/abio.1996.0034
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
High-resolution capillary gel electrophoresis was used for the separation of oligosaccharides labeled with a novel fluorophore 1-aminopyrene-3,6,8-trisulfonate (APTS) at the reducing termini by reductive amination. The APTS-saccharide adducts were detected by laser-induced fluorescence with excitation by the 488-nm Ar-ion laser and a 520-nm emission filter. The stoichiometry of labeling is such that only one molecule of fluorophore is attached to each molecule of oligosaccharide. Derivatization parameters, such as labeling reagent concentration, labeling temperature, and time as well as the influence of reaction solvent are thoroughly discussed. Desialylation of several sialylated oligosaccharides with different structures and linkages due to the effects of labeling temperature and time are also addressed. Employing the optimized conditions suggested in this paper, fluorophore labeling efficiency greater than 97% was achieved with no significant loss of sialic acid residues. The fluorescently labeled oligosaccharides are then separated and quantified by capillary gel electrophoresis. Practical examples of low-level derivatization and high-resolution capillary gel electrophoresis separation of N-linked glycans of ribonuclease-B and fetuin are also shown. (C) 1996 Academic Press, Inc.
引用
收藏
页码:234 / 242
页数:9
相关论文
共 42 条
[1]  
BARKER R, 1982, METHOD ENZYMOL, V83, P58
[2]   ANALYSIS OF MONOSACCHARIDE AND OLIGOSACCHARIDE ISOMERS DERIVATIZED WITH 9-AMINOPYRENE-L,4,6-TRISULFONATE BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE [J].
CHEN, FTA ;
EVANGELISTA, RA .
ANALYTICAL BIOCHEMISTRY, 1995, 230 (02) :273-280
[3]   CAPILLARY ZONE ELECTROPHORESIS OF OLIGOSACCHARIDES DERIVATIZED WITH VARIOUS AMINONAPHTHALENE SULFONIC-ACIDS [J].
CHIESA, C ;
ONEILL, RA .
ELECTROPHORESIS, 1994, 15 (8-9) :1132-1140
[4]   CAPILLARY ZONE ELECTROPHORESIS OF MALTOOLIGOSACCHARIDES DERIVATIZED WITH 8-AMINONAPHTHALENE-1,3,6-TRISULFONIC ACID [J].
CHIESA, C ;
HORVATH, C .
JOURNAL OF CHROMATOGRAPHY, 1993, 645 (02) :337-352
[5]   DETERMINATION OF CARBOHYDRATES BY CAPILLARY ZONE ELECTROPHORESIS WITH AMPEROMETRIC DETECTION AT A COPPER MICROELECTRODE [J].
COLON, LA ;
DADOO, R ;
ZARE, RN .
ANALYTICAL CHEMISTRY, 1993, 65 (04) :476-481
[6]   CHARACTERIZATION OF 9-AMINOPYRENE-1,4,6-TRISULFONATE-DERIVATIZED SUGARS BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION [J].
EVANGELISTA, RA ;
LIU, MS ;
CHEN, FTA .
ANALYTICAL CHEMISTRY, 1995, 67 (13) :2239-2245
[7]  
FEIZI T, 1989, J BIOCH, V245, P1
[8]  
FUKUDA MN, 1989, BLOOD, V73, P84
[9]   CAPILLARY ZONE ELECTROPHORESIS OF P-AMINOBENZOIC ACID-DERIVATIVES OF ALDOSES, KETOSES AND URONIC-ACIDS [J].
GRILL, E ;
HUBER, C ;
OEFNER, P ;
VORNDRAN, A ;
BONN, G .
ELECTROPHORESIS, 1993, 14 (10) :1004-1010
[10]   CAPILLARY GEL-ELECTROPHORESIS SEPARATION OF HIGH-MANNOSE TYPE OLIGOSACCHARIDES DERIVATIZED BY 1-AMINOPYRENE-3,6,8-TRISULFONIC ACID [J].
GUTTMAN, A ;
PRITCHETT, T .
ELECTROPHORESIS, 1995, 16 (10) :1906-1911