摘要
Phenothiazine化合物如Thionine、Toluidine Blue與Methylene Blue具有黏附蛋白質的應用潛力，本實驗以電化學氧化聚合法以及偶氮化-去氮還原固定法將Thionine、Toluidine Blue以及Methylene Blue固定於導電玻璃 (ITO) 上，再藉由Flow Injection Analysis以及Chronoamperometry技巧，對這三種物質吸附葡萄糖氧化脢、儲鐵蛋白以及鮭魚精蛋白之能力進行比較。實驗顯示：Thionine最具應用潛力，當蛋白質被逐量注入時，蛋白質會被吸附於Thionine表面，導致其螢光逐漸減弱，與利用掃描探針式顯微鏡影像分技術 (AFM) 所得結果極吻合。雖然Chronoamperometry也具有類似性質，但其靈敏性較差，因此難以有效檢測蛋白質含量。根據上述實驗結果，我們證實Phenothiazine化合物具有作為蛋白質分子膠的應用潛力，而且漫射式螢光光譜分析法是檢測蛋白質含量的有效工具之一。
並列摘要
Phenothiazines, such as thionine chloride (TC), toluidine blue (TB) and methylene blue (MB) are effective molecular adhesives for protein immobilization. In the light of the potential of phenothiazines in this respect, we prepared phenothiazine-modified electrodes via anodic polymerization and diazotization-reduction processes. With the help of diffuse-fluorescence spectroscopy (DFS) and flow injection analysis (FIA), we compared the biocompatibility of phenothiazines with glucose oxidase (GOx), ferritin (FT) and salmon testes (DNA). According to DFS and conductive-mode atomic force microscopy (C-AFM), we confirmed that TC is an effective molecular glue for GOx, compared to its structural analogues, TB and MB. When GOx was injected over the TC-modified electrodes, it could adsorb on TC and causing the fluorescence to decrease. We also compared DFS with other techniques in GOx analysis, such as chronoamperometry, DFS stood out as the most sensitive means. Based on these results, we conclude that phenothiazines are useful glues for protein immobilization, and diffuse-fluorescence spectroscopy is an effective tool for protein analysis.
參考文獻
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延伸閱讀
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