Bifunctional labeling reagent for oligosaccharides to incorporate both chromophore and biotin groups

被引:52
作者
Shinohara, Y [1 ]
Sota, H [1 ]
Gotoh, M [1 ]
Hasebe, M [1 ]
Tosu, M [1 ]
Nakao, J [1 ]
Hasegawa, Y [1 ]
Shiga, M [1 ]
机构
[1] DOJINDO LABS,MASHIKI,KUMAMOTO 86122,JAPAN
关键词
D O I
10.1021/ac960004f
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We have developed a convenient and effective method for biotinylation of oligosaccharides at their reducing ends. A novel biotin hydrazide having a phenyl group produced the biotin adduct of N-acetyllactosamine (LacNAc) by simple incubation at 90 degrees C for 1 h. Although the biotin adduct was obtained as a mixture of several stereoisomers, one of the isomers, cyclic beta-glycoside, became predominant upon letting the reaction mixture stand in a weakly acidic state (pH 3.5). This conversion may be very advantageous for functional analysis of oligosaccharides because natural N-linked oligosaccharides exist in the cyclic beta form, The limit of detection of labeled LacNAc in reversed-phase chromatography was 330 fmol and showed good linearity in the range from 330 fmol to 261 pmol. When this procedure was applied to complex type and high mannose type N-linked oligosaccharides, the labeled oligosaccharides were easily detected and separated by reversed-phase, gel filtration, and anion exchange chromatographies. Furthermore, these labeled oligosaccharides were able to be immobilized onto the solid phase using avidin-biotin technology and were stable enough to allow the binding assay to be performed repeatedly and under the conditions for in situ exoglycosidase digestion. These results suggest that this derivatization technique might be useful for both separation and functional analysis of oligosaccharides.
引用
收藏
页码:2573 / 2579
页数:7
相关论文
共 26 条
[11]   SYNTHESIS OF 1-N-GLYCYL BETA-OLIGOSACCHARIDE DERIVATIVES - REACTIVITY OF LENS-CULINARIS LECTIN WITH A FLUORESCENT LABELED STREPTAVIDIN PSEUDOGLYCOPROTEIN AND IMMOBILIZED NEOGLYCOLIPID [J].
MANGER, ID ;
WONG, SYC ;
RADEMACHER, TW ;
DWEK, RA .
BIOCHEMISTRY, 1992, 31 (44) :10733-10740
[12]  
MARER NL, 1992, GLYCOBIOLOGY, V2, P49
[13]   PREPARATION OF A PHOTOACTIVATABLE FLUORESCENT DERIVATIVE OF LACTOSE AND ITS APPLICATION TO PHOTOAFFINITY-LABELING OF A CONGER EEL LECTIN [J].
MURAMOTO, K ;
YAMAUCHI, F ;
KAMIYA, H .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1994, 58 (06) :1013-1017
[14]   DETERMINATION OF RATE AND EQUILIBRIUM BINDING CONSTANTS FOR MACROMOLECULAR INTERACTIONS USING SURFACE-PLASMON RESONANCE - USE OF NONLINEAR LEAST-SQUARES ANALYSIS-METHODS [J].
OSHANNESSY, DJ ;
BRIGHAMBURKE, M ;
SONESON, KK ;
HENSLEY, P ;
BROOKS, I .
ANALYTICAL BIOCHEMISTRY, 1993, 212 (02) :457-468
[15]   HIGH-RESOLUTION H-1-NMR AND C-13-NMR SPECTRA OF D-GLUCOPYRANOSE, 2-ACETAMIDO-2-DEOXY-D-GLUCOPYRANOSE, AND RELATED COMPOUNDS IN AQUEOUS-MEDIA [J].
PERKINS, SJ ;
JOHNSON, LN ;
PHILLIPS, DC ;
DWEK, RA .
CARBOHYDRATE RESEARCH, 1977, 59 (01) :19-34
[16]  
PINCKARD RN, 1978, HDB EXPT IMMUNOLOGY, V1, pCH17
[17]   THE USE OF STREPTAVIDIN BIOTINYLGLYCANS AS A TOOL FOR CHARACTERIZATION OF OLIGOSACCHARIDE-BINDING SPECIFICITY OF LECTIN [J].
SHAO, MC .
ANALYTICAL BIOCHEMISTRY, 1992, 205 (01) :77-82
[18]  
SHINOHARA Y, 1995, J BIOCHEM-TOKYO, V117, P1076
[19]   KINETIC MEASUREMENT OF THE INTERACTION BETWEEN AN OLIGOSACCHARIDE AND LECTINS BY A BIOSENSOR BASED ON SURFACE-PLASMON RESONANCE [J].
SHINOHARA, Y ;
KIM, F ;
SHIMIZU, M ;
GOTO, M ;
TOSU, M ;
HASEGAWA, Y .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1994, 223 (01) :189-194
[20]   ADVANCES IN THE USE OF BIOTINYLATED DIAMINOPYRIDINE (BAP) AS A VERSATILE FLUORESCENT TAG FOR OLIGOSACCHARIDES [J].
TOOMRE, DK ;
VARKI, A .
GLYCOBIOLOGY, 1994, 4 (05) :653-663