| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1986: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1985: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Research Abstract |
1. Preparation of tritium-labeled oligosaccharides and monosaccharide alcohols.
The radioactive asparagine-linked oligosaccharides were prepared from human prothrombin, human IgG, human transferrin, hCG, bovine C1q, and bovine blood coagulation factors, prothrombin, factor X, and factor IX by hydrazinolysis followed by NaB <^3H_4> -reduction. The radioactive monosaccharide alcohols were also prepared by NaB <^3H_4> -reduction of monosaccharides contained in the sugar chains of glycoproteins.
2. Development of the methods for micro-determination of sugar chains by HPLC.
Thirty one HPLC columns prepared for the analysis of monosaccharides or oligosaccharides were tested for the separation of the radioactive carbohydrates obtained above, for the reproducibility of the separation, and for the quantitative elution of the carbohydrates from the column. The property of each column was analyzed by changing the column temperature, the flow rate, the elution buffer, etc. As the results, we are successful in the developments of the quantitative separations of monosaccharide alcohols mixture within 80 min, of acidic oligosaccharides within 60 min, of the neutral oligosaccharides up to 25 glucose units by their size within 75 min which is useful for the monosaccharide sequencing of the oligosaccharides, and of the oligosaccharides which have affinities to lectins within 90 min. These results will become a basis for the development of automatic analyzers for oligosaccharides in future.
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