Topoglidis E, Cass A E, Gilardi G, Sadeghi S, Beaumont N, Durrant J R
Department of Biochemistry, Imperial College of Science, Technology and Medicine, South Kensington, London SW7 2AY, U.K.
Anal Chem. 1998 Dec 1;70(23):5111-3. doi: 10.1021/ac980764l.
We have investigated the use of optically transparent, nanoporous TiO(2) films as substrates for protein immobilization. Immobilization on such films may be readily achieved from aqueous solutions at 4 °C. The nanoporous structure of the film greatly enhances the active surface area available for protein binding (by a factor of 150 for a 4-μm-thick film). We demonstrate that the redox state of immobilized cytochrome c may be modulated by the application of an electrical bias potential to the TiO(2) film and that the fluorescence yield of immobilized fluorophore-labeled maltose-binding protein may be used to monitor specifically maltose concentration. We conclude that nanoporous TiO(2) films may be useful both for basic studies of protein/electrode interactions and for the development of array-based bioanalytical devices employing both optical and electrochemical signal transduction methodologies.
我们研究了使用光学透明的纳米多孔TiO₂薄膜作为蛋白质固定的基质。在4°C下，可轻松从水溶液中将蛋白质固定在这种薄膜上。薄膜的纳米多孔结构极大地增加了可用于蛋白质结合的活性表面积（对于4μm厚的薄膜，增加了150倍）。我们证明，通过向TiO₂薄膜施加电偏压，可以调节固定化细胞色素c的氧化还原状态，并且固定化荧光团标记的麦芽糖结合蛋白的荧光产率可用于特异性监测麦芽糖浓度。我们得出结论，纳米多孔TiO₂薄膜对于蛋白质/电极相互作用的基础研究以及采用光学和电化学信号转导方法的基于阵列的生物分析设备的开发可能都有用。
