By Gerald W. Hart (auth.), Pierre Thibault, Susumu Honda (eds.)
Recent advancements in high-resolution separation recommendations according to capillary-scale chromatography and electrophoresis have reworked the research of unfastened and conjugated mono- and oligosaccharides. In Capillary Electrophoresis of Carbohydrates, hands-on specialists describe state-of-the-art suggestions in capillary electrophoresis (CE) for the research of complicated carbohydrates. Written in step by step aspect to make sure profitable experimental effects, those without problems reproducible protocols offer equipment for pattern instruction and research of mono- and oligosaccharides, glycoproteins, and glycoconjugates. assorted CE separation codecs, together with capillary sector electrophoresis (CZE), capillary isoelectric focusing (CIEF), capillary isotachophoresis (CITP), capillary gel electrophoresis (CGE), and micellar electrokinetic chromatography (MEKC), are offered for various varieties of functions, equivalent to the research of glycoforms, the separation of enantiomers, and the solution of oligosaccharides. The separation of local derivatized carbohydrates is gifted utilizing UV, fluorescence, and mass spectrometric detection. different purposes show using CE as an assay strategy for tracking glycosyltransferase job and for making a choice on the organization consistent of carbohydrate-protein interplay. an invaluable appendix describes the buildings of the main in general encountered carbohydrate residues and oligosaccharides from mammalian and bacterial origins. every one protocol comprises targeted details on reagents, equipment, and invaluable reviews and notes on methods.
well timed and hugely useful, Capillary Electrophoresis of Carbohydrates presents either beginner and skilled CE analysts with robust instruments for the profitable separation of complicated carbohydrates by means of capillary electrophoresis.
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Extra info for Capillary Electrophoresis of Carbohydrates
1 mg/mL of glycan mixture, 2 M HCOOH, 10% acetonitrile. The numbers above each peak indicate the degree of polymerization. 55 56 Suzuki et al. Fig. 11. Separation of PTMAA-derivatized maltooligosaccharides by CE-ESMS. (A) Total ion electropherogram, m/z 400–1300 and (B) Contour profile of m/z vs time (intensity of each peak is orthogonal to the plane). Conditions: 40 nL inj. 1 mg/mL of glycan mixture, 2 M HCOOH, 10% acetonitrile. The numbers beside each peak indicate the degree of polymerization.
Rev. Biochem. 62, 65–100. 7. , Wheeler, S. , Hunter, A. , Dwek, R. , and Harvey, D. J. (1997) Sequencing of N-linked oligosaccharides directly from protein gels: in-gel deglycosylation followed by matrix-assisted laser desorption/ionization mass spectrometry and normal-phase high-performance liquid chromatography. Anal. Biochem. 250, 82–101. 8. Rudd, P. , Guile, G. , Harvey, D. , and Dwek, R. A. (1997) Oligosaccharide sequencing technology. Nature 388, 205–207. 9. Rudd, P. , Morgan, B. , Wormald, M.
18. All chemistry has been completed by this step and the system is being washed before the next run. 0 mL). If it is left in this solution at elevated temperatures there is a possibility of desialylation of the released glycans and the sample should therefore be dried down as soon as possible after collection. Acknowledgments The authors would like to thank Dr. Terry Butters, Neil Murphy, and Christina Colominas of the Oxford GlycoBiology Institute, and Dr. Cathy Merry of the Cancer Research Campaign, Department of Medical Oncology, Christie Release of Glycans from Glycoproteins 39 Hospital, Manchester for their advice and constructive criticism of the manuscript.