| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Research Abstract |
To evaluate binding event between protein and its ligand, an electrode covered with a protein/biomaterial film was constructed. Chitin and collagen, which have a high biocompatibility, were used as a substitute of cell surface. The protein having molecular recognition function was immobilized on the basis of electrostatic interaction between the protonated functional group in the biomaterial and the negative charged protein.
Evaluation of streptavidin(pI 5.0)-biotin binding was attempted at an electrode covered with chitin film. When the electrode modified with streptavidin was incubated in a solution containing biotin labeled with an electroactive compound, the peak current of LB decreased by holding of the electrode active moiety in the binding site of streptavidin to biotin. In contrast, the electrode response of only label did not change. The result showed that the LB combined with streptavidin immobilized on chitin film surface.
Using collagen film, investigation of lectin-sugar interaction was carried out. The lectin-sugar binding relates to physiological process. Concanavalin A (Con A), which is one of lectins, recognizes mannose moiety. Therefore, mannose with an electroactive compound was prepared. Con A-labeled mannose binding at an electrode covered with Con A/collagen film made the peak current of labeled mannose decrease. On the other hand, the decrease of peak current of labeled galactose and labeled glucose was not observed. Thus, Con A was immobilized on the chitin film surface, and monitoring of the binding was achieved.
Consequently, the biopolymers are powerful as a reaction field for a protein-ligand binding. This concept would be applied to design of sensor that evaluates the protein-lignd binding in vivo.
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