キチン−キトサン膜で被覆された電極を用いるタンパク質−リガンド間結合の電気化学的評価

概要

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An electrode covered with chitin-chitosan composite film was constructed to monitor binding between protein and ligand. Chitin, which is one of the extracellular matrixes, was dissolved in an organic solvent of N,N-dimethylacetoamide and 1-methyl-2-pyrrolidone. Because chitosan powder remained undissolved, it was considered to be a reaction field of biomolecule interactions. To examine whether the film was powerful, avidin and biotin were selected as a model of a protein and its ligand. Biotin moieties were introduced to active amino groups of the chitosan with a biotinylation reagent. To evaluate the binding on the film, biotin labeled with an electroactive daunomycin was used as a probe. As a result, an oxidation peak of the labeled biotin decreased. This was because the labeled biotin was held in the binding sites of avidin for biotin not used to be immobilized on a biotin-modified chitosan particle. Consequently, the binding could be monitored on the chitin-chitosan film with high biocompatibility. The procedure becomes a new method to evaluate binding between protein and ligand.