Humana Capillary Electrophoresis of Carbohydrates: 213 (Methods in Molecular Biology, 213)

Glycoconjugates such as glycoproteins and glycolipids play important roles in cell-cell interaction events, including development, differentiation, m- phogenesis, fertilization, inflammation, and metastasis. A number of reports have documented the association of unique oligosaccharide sequences to p- tein targeting and folding, and in mechanisms of infection, inflammation, and immunity. For glycoproteins, these glycan appendages are the result of extensive co- or post-translational modifications of the nascent proteins in the endoplasmic reticulum and in the Golgi apparatus. Although nucleic acids and proteins are copied from a template in a repeated series of identical steps using the same enzymes, complex carbohydrates are formed by the sequential actions of cellular glycosyltransferases that specifically recognize unique s- strates. The molecular biology of these transferases and other carbohydra- modifying enzymes is providing important insights on oligosaccharide recognition events. While it is acknowledged that the definition of the protein complement of cells and tissues (the so-called proteome) remains an enormous task in this postgenomic era, the characterization of all glycans produced by individual organisms (referred to as the glycome) presents an equally imp- tant challenge. This task is further complicated by the fact that oligosacc- rides cannot presently be cloned. These complex carbohydrates exist in a staggering diversity of structures, linkages, and branching, thus providing an exquisite molecular repertoire for cellular interactions. Review "...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 From the Back Cover 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 Electropho