Cytochrome P450 modified polycrystalline indium tin oxide film as a drug metabolizing electrochemical biosensor with a simple configuration.
Cytochrome P450 modified polycrystalline indium tin oxide film as a drug metabolizing electrochemical biosensor with a simple configuration.
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DOI：
10.1021/ac402661w
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发表时间：
2013-10
影响因子：
7.4
通讯作者：
K. Yoshioka;Dai Kato;Tomoyuki Kamata;O. Niwa
中科院分区：
文献类型：
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作者：
K. Yoshioka;Dai Kato;Tomoyuki Kamata;O. Niwa
细胞色素P450（CYP450）蛋白生物催化电极的研制是建立简单的药物代谢生物传感器或药物抑制剂筛选装置的关键技术。我们已经成功地检测到直接电子转移（DET）从人的电子层或吸附在裸铟锡氧化物（ITO）膜电极没有任何修饰层的电子微粒体，并将其应用于药物代谢评价。我们比较了两种ITO薄膜的电催化性能与不同的表面纳米结构（多晶或非晶）。在多晶ITO膜上的电子转移速率比在非晶ITO膜上的电子转移速率高15倍。多晶ITO膜是一个合适的电极吸附的蛋白质，同时保持有效的DET和酶的活性，可能是因为其较大的表面积和带负电荷的表面。多晶ITO膜电极处的氧还原电流增加了3至4倍，特别是与药物（睾酮和奎尼丁）的氧化作用相结合。相反，氧还原电流完全消失的抑制剂（酮康唑）的存在下。同样的结果也可以从具有足够清晰反应的线粒体微粒体中获得。这些结果表明，修饰的多晶ITO电极提供了一个简单的配置电化学评价药物代谢的活性的潜力。
The development of a biocatalytic electrode consisting of cytochrome P450 (CYP) proteins would be a key technology with which to establish simple drug metabolizing biosensors or screening devices for drug inhibitors. We have successfully detected the direct electron transfer (DET) from a human CYP layer or a CYP microsome adsorbed on a bare indium tin oxide (ITO) film electrode without any modification layers and applied it to drug metabolism evaluation. We compared the electrocatalytic properties of the two ITO films with different surface nanostructures (polycrystalline or amorphous). CYP on polycrystalline ITO film enhanced the electron transfer rate of oxygen reduction about fifteen times more than with amorphous film. The polycrystalline ITO film was a suitable electrode for the adsorption of CYP proteins while maintaining efficient DET and enzymatic activity, probably because of its larger surface area and negatively charged surface. The oxygen reduction current at the polycrystalline ITO film electrodes had increased 3- to 4-fold, specifically coupled with the oxidation of drugs (testosterone and quinidine) by the monooxygenase activity of CYP. In contrast, the oxygen reduction current completely disappeared in the presence of the CYP inhibitor (ketoconazole). Similar results could be obtained from the CYP microsome with sufficiently clear responses. These results indicate that the CYP modified polycrystalline ITO electrode offers the potential for electrochemically evaluating CYP activity for drug metabolism with a simple configuration.