In this 5-day course, participants will learn basic techniques for the isolation and characterization of oligosaccharides via a combination of hands-on work, lectures and demonstrations. Glycosphingolipids will be separated by thin-layer chromatography and detection protocols will be discussed. The use of lectin blotting techniques for characterization of oligosaccharide structural features will be covered. Other topics to be included are release and structural characterization of N- and O-linked glycans, and monosaccharide composition analysis. In addition to this course, modules on using mass spectrometry, computer modeling or NMR spectroscopy for analysis of glycoconjugates will be included. Experience with basic biochemical techniques is a prerequisite for participation.
The course will feature hands-on laboratory work and will include demonstrations and lectures. A lab manual including selected analytical techniques and references will be provided. The cost of registration (including lunch) is $500 per course for individuals from nonprofit institutions; $1100 for others. The cost of travel and lodging is not included.
Sample of NIH 2015 Schedule
On Friday of this course, participants can choose one of the following three modules: Mass Spectrometric (MS) Analysis of Glycoconjugates; Computational Simulations of Carbohydrate Conformation; and NMR of Carbohydrates. The course participants should indicate on the course application form which module they would be interested in taking.
Mass Spectrometric (MS) Analysis of Glycoconjugates: The use of mass spectrometry for the characterization of glycoconjugates will be discussed. Topics in this area will include identifying occupied glycosylation sites by LC-MS and sequencing glycoprotein glycans with sequential exoglycosidase digestions followed by MS. Laboratory demonstrations will also be performed on these two topics.
Computational Simulations of Carbohydrate Conformation: The course will present a discussion of the conformational preferences of oligosaccharides. Focus will then shift to the prediction and validation of oligosaccharide structures produced computationally with molecular dynamics simulations with the AMBER/GLYCAM force field.
NMR of Carbohydrates: This module will be an introduction to the application of NMR spectroscopy of polysaccharides and glycoconjugates covering a) essential features of NMR spectra, b) protocols used to solve primary structures, iii) applications and examples from the current literature, and c) resources for information. Our four NMR instruments (300-, 500-, 600- and 800 MHz) will be used for demonstrations during this module.