糖蛋白質の糖鎖機能研究のための試料調製

抄録

Glycan analysis by mass spectrometry (MS) is an emerging field in glyco-proteomics, poised to meet the technical demand for elucidation of the structural complexity and functions of the glycan components of molecules. Considering the divergence of the mass spectrometric methods employed for glycan analysis, it is necessary to establish sample preparation and technical standards.
In the present study of the HUPO HGPI (Human Proteome Organization Human Disease Glycomics/ Proteome Initiative, http://www.hgpi.jp/), the same samples of transferrin (Tf) and immunoglobulin G (IgG), both of which have N-glycans, were analyzed for their glycan structures and their relative abundances in 20 laboratories, and the chromatographic and MS analysis results were reported.
The blood samples were obtained from three healthy Japanese donors with the permission from the Medical Ethics Committee of Osaka University Graduate School of Medicine. Tf and IgG were purified from individual serum by immunoaffinity with rabbit polyclonal antibody against human Tf, and by protein G affinity chromatography, respectively, and then lyophilized. The purity of distributed samples was validated by SDS-PAGE under reducing conditions. The stability of these materials during transport was guaranteed by test incubation, during which neither degradation nor modification of glycan moieties was observed after one week of storage at 37 degrees centigrade.
Glycomic and glyco-proteomic methodologies of the type assessed in this study, have the potential to provide large volumes of quantitative data for diagnostics and, more broadly, for underpinning functional glycomics and systems biology research.
A part of this work was supported by the 21st century COE Program of Osaka University from the Japan Society for the Promotion of Science (JSPS) and by the JSPS Core-to-Core program.
REFERENCE
Wada Y, et al. Glycobiology. 2007, 17:411-22.