Group Leader:
Dr. Jean-Robert Brisson
(613) 990-3244 /
Jean-Robert.Brisson@nrc.gc.ca

Business Contact:
Scott Ferguson,
(613) 990-5948 /
Scott.Ferguson@nrc.gc.ca

Group Focus:
The mandate of the Glycoanalysis group is the development of methods for determining the structure and interactions of carbohydrates in microbial virulence and in neurodegenerative disease. Carbohydrates and their associated metabolic and carbohydrate processing enzymes critical for virulence expression are identified and exploited as targets for drug discovery, glyco-engineering, vaccine design and detection and diagnosis of diseases. Carbohydrate antigens and their processing enzymes are identified as targets for prevention and treatment of neuronal degeneration or improvement of neuronal adaptation and regeneration. We are also involved in the identification of molecular targets for the development of vaccines and immunological therapies to alleviate infectious and neurodegenerative diseases and cancer. Hence, our group interacts with scientists in the Eukaryotic Glycobiology, Genomics and Proteomics, Glycosyltransferases and Neuroglycomics, Glycosidases, Antibody Engineering, Infection and Immunity, Immnumodulation, Antibody Engineering, and Molecular Pathogenesis groups at IBS.

The Team:
Dr. Jean-Robert Brisson, Nam Huan Khieu, Dr. David McNally, Dr. Harold Jarrell, Dr. Scott Houliston, Dr. Evgeny Vinogradov, Dr. Jianjun Li, Kenneth Chan, Jacek Stupak, Dr. Monika Dzieciatkowska, Dr. Xin Liu

Ongoing Research Activities and Projects:

1. Mass spectrometry and NMR spectral strategies for the structural analysis of bacterial glycoconjugates (i.e., lipopolysaccharides, capsular polysaccharides, glycoproteins).
2. Development of methods for the purification of glycoconjugates and highly labile bacterial carbohydrates.
3. Development of techniques for quantitative analysis of glycans on glycoproteins for prokaryotic and eukaryotic systems.
4. A capillary electrophoresis separation system for electrospray mass spectrometry.
5. Identification of trace level of glycans by MS and cold-probe NMR technologies for metabolomics and eukaryotic glycobiolgy applications.
6. Study of the glycan component of the lipo-oligosaccharide in the pathogenic bacteria C. jejuni and H. influenzae, which may be involved in the onset of Guillan-Barré syndrome.
7. High Resolution Magic Angle Spinning (HR-MAS) NMR spectral characterization of cell surface glycans on intact bacterial cells.
8. Proton NMR metabolomic analysis of biofluids.
9. Pulse sequence design for improved glycan structural analysis and the study of carbohydrate-protein interactions.
10. Solid-state NMR structural analysis of complex cellulose materials in processed hemp fibers.
11. Molecular basis of protein-carbohydrate interactions by NMR and molecular modeling.

Partnering Opportunities:
Service contracts, joint R&D projects and technical exchanges are on-going activities of the Glycoanalysis group.

Expertise and Available Technologies:

1. Mass Spectrometry Facility
   • The AB/MDS Sciex 4000 QTRAP is a hybrid instrument that combines the best features of triple quadrupole and ion trap mass spectrometers. The instrument has significantly greater sensitivity and resolving power compared to conventional triple quadrupole mass spectrometers thanks to the substitution of the third quadrupole with a linear ion trap. The Q-Trap is also capable of MS3 scanning (multiple stages of tandem mass spectrometry) which is essential for analyzing complex glycans and protein post translational modifications. This instrument retains all of the important scanning features of triple quadrupoles (i.e., precursor ion scanning, neutral loss scanning, etc.) which can be used to selectively detect analytes with specific features (i.e., sialylation, phosphorylation, etc.) and will facilitate in vivo analyses.
   • The AB/MDS Sciex API 3000 is a research grade triple quadrupole mass spectrometer. By monitoring diagnostic ions, we can quickly monitor and deduce modifications in a protein. Coupled with capillary electrophoresis, it is the main instrument used for routine carbohydrates including LPS and surface antigens.
   • The AB/MDS Sciex QStar couples the rugged front end of the API with a very sensitive and high resolution Time-Of-Flight (TOF) spectrometer. The result is a high resolution MS/MS instrument for structural analysis and protein sequencing.
   • The Varian Saturn 2000 GC/MS, a "Walk Up" instrument, is primarily used in sugar composition analysis and sugar linkage analysis.
   • The AB Voyager Elite MALDI TOF, the "work horse" of the group, is used as a "service instrument" for screening peptide synthesis, for checking the purity of extracted proteins or carbohydrates, and to characterize polymers.
   • More in depth and high-throughput analysis is performed using the QTOF 2, QTOF Ultima and M@ADI by Waters in the Genomics and Proteomics group.
2. Purification of glycans
   • Two HPLC machines, used for Dionex-type separations, reverse phase and ion-exchange separations
3. NMR Facility
   • High resolution Varian NMR spectroscopy at 600, 500, 400, and 200 MHz with solid state capabilities for the 400 MHz spectrometer.
   • (1H, 13C, 15N, 2H) cryogenically cooled probe (cold-probe) for optimal sensitivity on the 4 channel 600 MHz spectrometer.
   • (1H, 19F, X) MAS solid state capabilities on the 4 channel 400 MHz spectrometer.
   • (1H, 31P, X), (1H, X) and broadband probes (X, 1H) for biological samples.
   • Nano-probe for combinatorial chemistry.
   • HR-MAS for cellular domains (bacterial cells, metabolites, cancer cells).
   • Remote operation of spectrometers.
4. Molecular modeling
   • 'InsightII Accelrys' and Sybyl software packages for general molecular modeling
   • Affinity and Autodock programs for auto-docking of ligand to proteins
   • Amber software for statistical analysis and molecular dynamics simulation of oligosaccharides and polysaccharide conformational analysis
   • Gaussian and NWChem programs for electronic structure calculation
   • SGI, SUN and Linux cluster hardware are used for the above modeling programs.