Glycoconjugate Journal

, Volume 16, Issue 6, pp 271–281 | Cite as

Preparation and isolation of neoglycoconjugates using biotin-streptavidin complexes

  • B. Kuberan
  • N.S. Gunay
  • J.S. Dordick
  • R.J. Linhardt

Abstract

Glycoproteins commercially available in multi-gram quantities, were used to prepare milligram amounts of neoglycoproteins. The glycoproteins bromelain and bovine γ-globulin were proteolyzed to obtain glycopeptides or converted to a mixture of glycans through hydrazinolysis. The glycan mixture was structurally simplified by carbohydrate remodeling using exoglycosidases. Glycopeptides were biotinylated using N-hydroxysuccinimide activated-long chain biotin while glycoprotein-derived glycans were first reductively aminated with ammonium bicarbonate and then biotinylated. The resulting biotinylated carbohydrates were structurally characterized and then bound to streptavidin to afford neoglycoproteins. The peptidoglycan component of raw, unbleached heparin (an intermediate in the manufacture of heparin) was similarly biotinylated and bound to streptavidin to obtain milligram amounts of a heparin neoproteoglycan. The neoglycoconjugates prepared contain well defined glycan chains at specific locations on the streptavidin core and should be useful for the study of protein-carbohydrate interactions and affinity separations.

neoglycoconjugates neoglycoprotein neoproteoglycan biotin streptavidin heparin bromelain γ-globulin MWCO, molecular weight cut-off ANTS, 8-amino-1,3,6-napthalenetrisulfonate DMSO,dimethyl sulfoxide ESI,electrospray ionization NHS,N-hydroxysuccinimide LC,long chain CE,capillary electrophoresis PAGE,polyacryamide gel electrophoresis SDS,sodium dodecylsulfate BSA,bovine serum albumin SAX,strong-anion exchange 

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Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • B. Kuberan
    • 1
  • N.S. Gunay
    • 1
  • J.S. Dordick
    • 2
  • R.J. Linhardt
    • 1
  1. 1.Division of Medicinal and Natural Products Chemistry and Department of Chemical and Biochemical EngineeringUniversity of IowaIowa CityUSA
  2. 2.Department of Chemical EngineeringRensselaer Polytechnic InstituteTroyUSA

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