Abstract
The separation of highly diverse mixtures of glycoprotein glycans is often one of the major problems arising during carbohydrate structure analysis of glycoproteins. In the course of our studies on viral glycoproteins, we have established a fractionation procedure allowing the isolation of individual oligosaccharides at microscale. In order to facilitate the detection of glycoproteins and corresponding glycans during isolation and purification, they were biosynthetically labeled in their carbohydrate moieties by cultivation of virus-infected cells in the presence of [2-3H]Man or [6-3H]GlcN. The separation procedure used is based on sequential liberation of the different types of sugar chains present and their subsequent fractionation according to charge, size, and/or isomeric structure by a combination of different chromatographic techniques.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Tarentino, A. L., Gomez, C. A., and Plummer, T. H., Jr. (1985) Deglyco-sylation of asparagine-linked glycans by peptide:N-glycosidase F. Biochemistry 24, 4665–4671.
Schlüter, M., Linder, D., and Geyer, R. (1985) Isolation of glycopeptides containing individual glycosylation sites of Friend murine leukemia virus glycoprotein: Studies on glycosylation by methylation analysis. Carbohydr. Res. 138,305–314.
Pfeiffer, G., Geyer, H., Kalsner, I., Wendorf, P., and Geyer, R. (1990) Separation of glycoprotein-N-glycans by high-pH anion-exchange chromatography. Biomed. Chromatogr. 4,193–199.
Van Pelt, J., Damm, J. B. L., Kamerling, J. P., and Vliegenthart, J. F. G. (1987) Separation of sialyloligosaccharides by medium pressure anion-exchange chromatography on Mono Q. Carbohydr. Res. 169, 43–51.
Townsend, R. R., Hardy, M. R., Cumming, D. A., Carver, J. P., and Bendiak, B. (1989) Separation of branched sialylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection. Anal. Biochem. 182, 1–8.
Townsend, R. R., Hardy, M. R., Hindsgaul, O., and Lee, Y. C. (1988) High-performance anion-exchange chromatography of oligosaccharides using pellicular resins and pulsed amperometric detection. Anal. Biochem. 174, 459–470.
Van Pelt, J., Van Kuik, A. J., Kamerling, J. P., Vliegenthart, J. F. G., Van Diggelen, O. P., and Galjaard, H. (1988) Storage of sialic acid-containing carbohydrates in the placenta of a human galactosialidosis fetus. Isolation and structural characterization of 16 sialyloligosaccharides. Eur.J. Biochem. 177, 327–338.
Lamblin, G., Boersma, A., Lhermitte, M., Roussel, P., Mutsaers, H. G. M., Van Halbeek, H., and Vliegenthart, F. G. (1984) Further characterization, by a combined high-performance liquid chromatography/1H-NMR approach, of the heterogeneity displayed by the neutral carbohydrate chains of human bronchial mucins. Eur.J. Biochem. 143, 227–236.
Hardy, M. R. and Townsend, R. R. (1989) Separation of fucosylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection. Carbohydr. Res. 188, 1–8.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1993 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Geyer, R., Geyer, H. (1993). Isolation and Fractionation of Glycoprotein Glycans in Small Amounts. In: Hounsell, E.F. (eds) Glycoprotein Analysis in Biomedicine. Methods in Molecular Biology, vol 14. Humana Press. https://doi.org/10.1385/0-89603-226-4:131
Download citation
DOI: https://doi.org/10.1385/0-89603-226-4:131
Publisher Name: Humana Press
Print ISBN: 978-0-89603-226-2
Online ISBN: 978-1-59259-501-3
eBook Packages: Springer Protocols