Hands-On Training
The Complex Carbohydrate Research Center training includes undergraduate and graduate students, postdoctoral research associates, and visiting scientists from the United States and around the world. CCRC provides Hands-on Training in principles, methods, and analytical techniques used to study complex carbohydrates.
Short-term trainees may enroll in one or more of the four glycobiology training courses the CCRC offers every summer for scientists in academic, governmental, or industrial laboratories.
These varied hands-on trainings in glycobiology are supported in part by fees from participants and by funds from the US Department of Energy and the US National Institutes of Health.
Course 1 : Techniques for Characterization of Carbohydrate Structure of Plant and Microbial Polysaccharides - Polysaccharide Course (August 3-7, 2020).
This 5-day training course will cover the techniques of glycosyl residue (composition by alditol acetate and methanolysis) and glycosyl linkage (methylation) analyses using gas-liquid chromatography/mass spectrometry. Monosaccharide and oligosaccharide analysis by multiple methodologies include: high pH anion-exchange chromatography (HPAEC), techniques in purification and isolation of polysaccharides, and demonstrations and coverage of lignin analysis by chemical methods and Pyrolysis Molecular beam mass spectrometry (Py-MBMS) and Pyrolysis GC-MS. Participants will perform multiple hands-on analyses of purified oligosaccharides. Lectures and demonstrations will cover other techniques for structural analysis. In addition to this course, a choice of modules either on using mass spectrometry, computer modeling, or NMR spectroscopy for analysis of glycoconjuagtes 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.
Module Information
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.
Course 2 : Separation and Characterization of Glycoprotein and Glycolipid Oligosaccharides - Glycoprotein Course (August 10-14, 2020).
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.
Module Information
On Friday of this training 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.
Course 3 : Analytical Techniques for Structural Analysis of Glycosaminoglycans (GAGs). (August 17-20, 2020).
In this 4-day training course, participants will learn basic techniques for the structural characterization of glycosaminoglycans (GAGs), which will include depolymerization of the GAG polysaccharides to its disaccharide constituents using appropriate enzymes. The disaccharides will be separated by ion exchange chromatography and the products will be detected by U.V. The disaccharides will also be derivatized with 2-cyanoacetamide and will be detected with fluorescent detection. In a separate experiment, the glycosaminoglycans will be hydrolyzed by nitrous acid and the products will be analyzed. Mass spectrometry techniques for analysis of di- and oligosaccharides derived from glycosaminoglycans will also be covered. 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 $600 per course for individuals from nonprofit institutions; $1400 for others. The cost of travel and lodging is not included.
Course 4 : Glycomics, Glycoproteomics and Bioinformatics Tools for Data Interpretation of Mass Spectrometry of Glycoproteins (October 12-16, 2020).
The mass spectrometry and bioinformatics training course will focus on the analysis and data interpretation of glycoproteins by mass spectrometric techniques. MS component will include lectures on mapping the glycosylation sites in glycoproteins; determining the composition, sequencing, and branching points of N and O-linked oligosaccharides, and MS procedures used in these analyses. Demonstrations will include matrix-assisted laser desorption (MALDI), electrospray ionization (ES/MS), and tandem mass spectrometry (MS/MS). Experience with MS is beneficial but is not required.
The Bioinformatics component concentrates on the interpretation of mass spectrometric data and especially the application of bioinformatics tools for this purpose. The course is taught by Dr. Rene Ranzinger and the bioinformatics team at the CCRC with special lectures from Drs. Azadi and Archer-Hartmann. The course included lectures and hands-on exercises of:
- Introduction to different domain databases (Glycomics and GlycoProteomics) and how they can be used to supplement analytical workflows.
- Introduction in the interpretation of mass spectrometric data
- A general overview of existing mass spectrometry software tools for glycomics and glycoproteomics data
- Training in the interpretation of glycomics data using GlycoWorkbench and GRITS Toolbox
- Training in the interpretation of glycoproteomics data using Byonic™/Protein Discoverer ™
- Standards for reporting data and depositing data (e.g. MIRAGE, MIAPE)
A course manual including selected analytical techniques and references will be provided. The cost of registration (including lunch) is $500 per course for individuals from non-profit institutions and $1100 for others. The cost of travel and lodging is not included.