Rapid and simple preparation of N-linked oligosaccharides by cellulose-column chromatography

Citation
Y. Shimizu et al., Rapid and simple preparation of N-linked oligosaccharides by cellulose-column chromatography, CARBOHY RES, 332(4), 2001, pp. 381-388
Citations number
14
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Agricultural Chemistry","Chemistry & Analysis","Organic Chemistry/Polymer Science
Journal title
CARBOHYDRATE RESEARCH
ISSN journal
0008-6215 → ACNP
Volume
332
Issue
4
Year of publication
2001
Pages
381 - 388
Database
ISI
SICI code
0008-6215(20010615)332:4<381:RASPON>2.0.ZU;2-7
Abstract
As a means of preparing N-linked oligosaccharides from hydrazinolysates of glycoproteins in a rapid and simple manner, a method has been developed usi ng cellulose-column chromatography. Hydrazinolysates of human IgG, containi ng a series of biantennary complex type oligosaccharides, were applied to a cellulose column equilibrated with (4:1:1, v/v) 1-butanol-ethanol-water. T he N-linked oligosaccharides were eluted with (1:1, v/v) ethanol-water, and analyzed by NPLC in combination with sequential glycosidase digestion. The oligosaccharides, with or without sialic acid, were quantitatively recover ed in the fraction eluted with (1:1, v/v) ethanol-water without UV-detectab le contamination by impurities derived from protein or the cellulose. Other types of N-linked oligosaccharides of al-acid glycoprotein (tetraantennary complex-type), ovalbumin (hybrid-type), and ribonuclease B thigh mannose-t ype) were also quantitatively prepared from the hydrazinolysates by elution of the cellulose column with (1:1, v/v) ethanol-water and these had as hig h a quality as those prepared by conventional paper chromatography. (C) 200 1 Elsevier Science Ltd. All rights reserved.