6533b7d3fe1ef96bd1261251
RESEARCH PRODUCT
Analytical technique for studying the structure of glycoprotein N-glycans
Christine SchaumannFranz OeschKlaus K. UngerRaimund J. Weisersubject
Gel electrophoresischemistry.chemical_classificationGlycanChromatographybiologyImmobilized enzymeChemistryOrganic ChemistryGeneral MedicineBiochemistryWheat germ agglutininAnalytical Chemistrychemistry.chemical_compoundBiochemistryAffinity chromatographyConcanavalin Abiology.proteinGlycoproteinDerivatizationdescription
Abstract The aim of this study was to develop an analytical strategy for the structural analysis of glycoprotein N-glycans by combining several sensitive methods without any elaborate equipment. The following consecutive steps were optimized and applied: (1) immobilization of glycopeptide N-glycosidase F (EC 3.2.2.18) on several polymeric and a silica support, the latter giving a maximum binding capacity of 11.3% of starting activity; (2) lectin affinity chromatography was miniaturized using Mobitec columns of volume 200 μl; the binding capacity of glycoproteins on concanavalin A and wheat germ agglutinin columns was in the range 0.5–1 μg; (3) N-linked oligosaccharides were isolated from commercial glycoproteins and from contactinhibin, a glycoprotein of Mr 60 000–70 000. After derivatization with 8-aminonapthalene-1,3,6-trisulphonic acid, they were separated by high-resolution electrophoresis.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1993-08-01 | Journal of Chromatography A |