title: Protecting group-free immobilization of glycans for affinity chromatography using glycosylsulfonohydrazide donors.
abstract: A variety of applications in glycobiology exploit affinity chromatography through the immobilization of glycans to a solid support. Although several strategies are known, they may provide certain advantages or disadvantages in how the sugar is attached to the affinity matrix. Additionally, the products of some methods may be hard to characterize chemically due to non-specific reactions. The lack of specificity in standard immobilization reactions makes affinity chromatography with expensive oligosaccharides challenging. As a result, methods for specific and efficient immobilization of oligosaccharides remain of interest. Herein, we present a method for the immobilization of saccharides using N'-glycosylsulfonohydrazide (GSH) carbohydrate donors. We have compared GSH immobilization to known strategies, including the use of divinyl sulfone (DVS) and cyanuric chloride (CC), for the generation of affinity matrices. We compared immobilization methods by determining their immobilization efficiency, based on a comparison of the mass of immobilized carbohydrate and the concentration of active binding sites (determined using lectins). Our results indicate that immobilization using GSH donors can provide comparable amounts of carbohydrate epitopes on solid support while consuming almost half of the material required for DVS immobilization. The lectin binding capacity observed for these two methods suggests that GSH immobilization is more efficient. We propose that this method of oligosaccharide immobilization will be an important tool for glycobiologists working with precious glycan samples purified from biological sources.
authors.affiliation: Alberta Glycomics Centre, Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Hernandez Armada
authors.affiliation: Alberta Glycomics Centre, Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
authors.forename: Jobette T
authors.initials: JT
authors.lastname: Santos
authors.affiliation: Alberta Glycomics Centre, Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
authors.forename: Michele R
authors.initials: MR
authors.lastname: Richards
authors.affiliation: Alberta Glycomics Centre, Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada. Electronic address: ccairo@ualberta.ca.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Cairo
mesh_terms: D001665:Binding Sites; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D054794:Glycomics / Q000295:instrumentation* / Q000379:methods; D006834:Hydrazines / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry*; D013450:Sulfones / Q000737:chemistry; D014227:Triazines / Q000737:chemistry
chemical_list: D006834:Hydrazines; D009844:Oligosaccharides; D037121:Plant Lectins; D013450:Sulfones; D014227:Triazines; C009873:divinyl sulfone; C019309:cyanuric chloride; D012685:Sepharose
keywords: Affinity chromatography; Divinyl sulfone; Glycosylsulfonohydrazide; Immobilization; Lectin; Protecting group-free