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Recent developments in high-resolution separation techniques based on capillary-scale chromatography and electrophoresis have transformed the analysis of free and conjugated mono- and oligosaccharides. In Capillary Electrophoresis of Carbohydrates, hands-on experts describe cutting-edge techniques in capillary electrophoresis (CE) for the analysis of complex carbohydrates. Written in step-by-step detail to ensure successful experimental results, these readily reproducible protocols provide methods for sample preparation and analysis of mono- and oligosaccharides, glycoproteins, and glycoconjugates. Different CE separation formats, including capillary zone electrophoresis (CZE), capillary isoelectric focusing (CIEF), capillary isotachophoresis (CITP), capillary gel electrophoresis (CGE), and micellar electrokinetic chromatography (MEKC), are presented for different types of applications, such as the analysis of glycoforms, the separation of enantiomers, and the resolution of oligosaccharides. The separation of native derivatized carbohydrates is presented using UV, fluorescence, and mass spectrometric detection. Two other applications demonstrate the use of CE as an assay method for monitoring glycosyltransferase activity and for determining the association constant of carbohydrate-protein interaction. A useful appendix describes the structures of the most commonly encountered carbohydrate residues and oligosaccharides from mammalian and bacterial origins. Each protocol contains detailed information on reagents, apparatus, and valuable comments and notes on methods.
Timely and highly practical, Capillary Electrophoresis of Carbohydrates provides both novice and experienced CE analysts with powerful tools for the successful separation of complex carbohydrates by capillary electrophoresis.
Table des matières
Structural and Functional Diversity of Glycoconjugates.- Sample Preparation Methods.- Chemical and Enzymatic Release of Glycans from Glycoproteins.- Derivatization of Carbohydrates.- Preparation of Oligosaccharides from Sulfated Glycosaminoglycans Using Bacterial Enzymes.- Analysis of Mono- and Oligosaccharides.- Monosaccharide Analysis by Capillary Electrophoresis.- Separation of Oligosaccharides by Capillary Electrophoresis Using Buffer Modifiers.- Characterization of Oligosaccharides from Starch, Dextran, Cellulose, and Glycoproteins by Capillary Electrophoresis.- Affinity Capillary Electrophoresis of Oligosaccharides.- Characterization of Glycosaminoglycans by Capillary Electrophoresis.- Analysis of Glycoconjugates and the Component Monosaccharides.- Analysis of 2-Aminoacridone-Derivatized Complex Oligosaccharides Using Micellar Electrokinetic Capillary Chromatography and Laser-Induced Fluorescence Detection.- Separation of Protein Glycoforms by Capillary Electrophoresis.- Separation of Glycoproteins by Capillary Isoelectric Focusing.- Glycoprotein Analysis by Capillary Zone Electrophoresis-Electrospray Mass Spectrometry.- Analysis of Bacterial Glycolipids by Capillary Electrophoresis-Electrospray Mass Spectrometry.- Specific Applications.- Capillary Electrophoresis as an Assay Method for Monitoring Glycosyltransferase Activity.- Determination of Association Constant of Carbohydrate-Protein Interaction.
Résumé
A collection of techniques for using Capillary Electrophoresis (CE) to analyze complex carbohydrates. It provides methods for sample preparation, analysis of mono- and oligosaccharides, glycoproteins, and glycoconjugates. It describes the structures of the carbohydrate residues and olgosaccharides from mammalian and bacterial origins.
Texte suppl.
"...the book will be very valuable for researchers carrying out carbohydrate research, or planning to do so, and contemplating the best analytical approaches." -Pharmaceutical Research
"...this book provides a great deal of useful practical detail for CE analysis of carbohydrates for the experienced as well as the newcomer to the field." -Biomedical Chromatography
"...the book is a nice compilation of protocols for the analysis of carbohydrates by CE...it does contain all the essential procedures and methods needed to solve many problems." - Chromatographia
"...this book comprehensively summarizes knowledge on carbohydrate analysis by means of capillary electrophoresis." - Biochemistry
Commentaire
"...the book will be very valuable for researchers carrying out carbohydrate research, or planning to do so, and contemplating the best analytical approaches." -Pharmaceutical Research
"...this book provides a great deal of useful practical detail for CE analysis of carbohydrates for the experienced as well as the newcomer to the field." -Biomedical Chromatography
"...the book is a nice compilation of protocols for the analysis of carbohydrates by CE...it does contain all the essential procedures and methods needed to solve many problems." - Chromatographia
"...this book comprehensively summarizes knowledge on carbohydrate analysis by means of capillary electrophoresis." - Biochemistry
