Development of methods to determine carbohydrate structure by Fourier transform mass spectrometry

seven 2-aminobenzamide (2ab)-derivatized oligosaccharides could be determined assmall as 10pmol by matrix-assisted laser desorption ionization (MALDI)-Fourier transform massspectrometry (FTMS) with a good signal-to-noise ratio. Interestinglyall ions from either high-mannose or comples -type sugar chains have shown spontaneously with theGIcNac-3AB by B-type cleavage. Additionally，loss of two hexoses was found from nonreducing terminal from bi-, tri-，tetra-antennary complex-type sugars，关于galactose residues on GIcNAcβ1-2Man arm may be deleted specifically. on the otherhand, Man D29 D2GIcNAc D22 D2-2AB released three hexoses from nonreducing termini，suggesting three Manα1-2 linkages may be fragile. 2ab -derivatized oligosaccharides were separated bythree lectin column chromatographies and then subjected to MALDI time-of-flight mass spectrometry（MALDI-TOF/MS） for structural characterization of the carbohydrates. the combination…sequential exoglyconsidase digestion and MALDI-TOF/MS greatly facilitatesmonosaccharide sequencing and is more feasible than size-exclusion column chromatography in terms ofthe time consumed and the laboriousness of the procedure. By this strategymicrosequencing of 2-3 pmol of oligosaccharide derivatives could be achieved. Furthermorespectra obtained by the post source decay (PSD) mode provide excellent sequence信息。相关intensities of metastable ions due to fragmentation at glycosidic linkages were differentamong linkage isomers of particular oligosaccharides.这些研究解释PSD分析possesses significant potential for the estimation of glycosidic linkage in carbohydrate结构。The quantitative nature of MALDI-TOF/MS in oligosaccharide analysis was of great use in Thecomparative study on normal and pathogenic prion proteins, PrP - D1c - D1 and PrP - D1Sc - D1，Our developed methods enable analysis of glycan mixtures quantitatively. Less

Molecular weight of seven 2-aminobenzamide (2AB)-derivatized oligosaccharides could be determined as small as 10 pmol by matrix-assisted laser desorption ionization (MALDI)-Fourier transform mass spectrometry (FTMS) with a good signal-to-noise ratio. Interestingly, all ions from either high-mannose or complex-type sugar chains have shown spontaneously with the release of GIcNac-3AB by B-type cleavage. Additionally, loss of two hexoses was found from nonreducing terminal from bi-, tri-, and tetra-antennary complex-type sugars, suggesting that galactose residues on GIcNAcβ1-2Man arm may be deleted specifically. On the other hand, ManィイD29ィエD2GIcNAcィイD22ィエD2-2AB released three hexoses from nonreducing termini, suggesting three Manα1-2 linkages may be fragile. 2AB-derivatized oligosaccharides were separated by three lectin column chromatographies and then subjected to MALDI time-of-flight mass spectrometry (MALDI-TOF/MS) for structural characterization of the carbohydrates. The combination … More of sequential exoglyconsidase digestion and MALDI-TOF/MS greatly facilitates the monosaccharide sequencing and is more feasible than size-exclusion column chromatography in terms of the time consumed and the laboriousness of the procedure. By this strategy, microsequencing of 2-3 pmol of oligosaccharide derivatives could be achieved. Furthermore, spectra obtained by the post source decay (PSD) mode provide excellent sequence information. The relative intensities of metastable ions due to fragmentation at glycosidic linkages were different among linkage isomers of particular oligosaccharides. These results demonstrate that PSD analysis possesses significant potential for the estimation of glycosidic linkage in carbohydrate structures. The quantitative nature of MALDI-TOF/MS in oligosaccharide analysis was of great use in the comparative study on normal and pathogenic prion proteins, PrPィイD1cィエD1 and PrPィイD1ScィエD1, respectively. Our developed methods enable analysis of glycan mixtures quantitatively. Less