This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Course: Analytical Techniques for Carbohydrate Structure Determination This course covered the techniques of glycosyl residue (composition) and glycosyl linkage (methylation) analyses using gas-liquid chromatography/mass spectrometry. Participants performed analyses of purified oligosaccharides, and a few brought their own samples. Lectures and demonstrations covered other techniques for structural analysis. In addition, a choice of modules either on using mass spectrometry or NMR spectroscopy for analysis of glycoconjugates was included (see descriptions below). Experience with basic biochemical techniques was a prerequisite for participation. Course: Separation and Characterization of Glycoconjugate Oligosaccharides Participants learned basic techniques for the isolation and characterization of oligosaccharides via a combination of hands-on work, lectures and demonstrations. Mixtures of oligosaccharides or glycopeptides derived from glycoproteins were resolved into individual components by chromatographic procedures such as size-exclusion, HPLC, lectin affinity chromatography, and/or polyacrylamide gel electrophoresis. Glycosphingolipids were separated by thin-layer chromatography, and detection protocols were discussed. The use of lectin blotting techniques for characterization of oligosaccharide structural features was covered. Other topics included were conjugation of oligosaccharides with fluorescent tags, capillary electrophoresis and monosaccharide composition analysis. In addition, modules on using mass spectrometry and/or NMR spectroscopy for analysis of glycoconjugates was included (see descriptions below). Experience with basic biochemical techniques was a prerequisite for participation. Additional Course Modules Mass Spectrometric (MS) Analysis of Glycoconjugates: The use of mass spectrometry for the characterization of glycoconjugates was discussed. Topics in this area included identifying occupied glycosylation sites by LC-MS and sequencing glycoprotein glycans with sequential exoglycosidase digestions followed by MS. Laboratory demonstrations were also be performed on these two topics.
Showing the most recent 10 out of 245 publications
