Wang Jianlong, Wang Fuan, Chen Hongjun, Liu Xiaohua, Dong Shaojun
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Changchun Jilin 130022, Graduate School of the Chinese Academy of Sciences, Beijing 100039, China.
Talanta. 2008 May 15;75(3):666-70. doi: 10.1016/j.talanta.2007.11.062. Epub 2007 Dec 7.
In this paper, electrochemical surface plasmon resonance (SPR) method was first used to detect enzymatic reaction in bilayer lipid membrane (BLM) based on immobilizing horseradish peroxidase (HRP) in the BLMs supported by the redox polyaniline (PAn) film. By SPR kinetic curve in situ monitoring the redox transformation of PAn film resulted from the reaction between HRP and PAn, the enzymatic reaction of HRP with H(2)O(2) was successfully analyzed by electrochemical SPR spectroscopy. The results show that this BLM supported on PAn film cannot only preserve the bioactivity of HRP immobilized in the membrane, but also provide a channel for the transfer of electrons between HRP and PAn on electrode surface. These characteristics enabled the development of SPR biosensor for sensitively detecting H(2)O(2). H(2)O(2) has been detected by electrochemical SPR spectroscopy in the concentration range of 5 x 10(-5)M to 2 x 10(-3)M. After each of detections, the SPR sensor surface was completely regenerated by electrochemically reducing the oxidized PAn to its reduced state. This method provides a novel route for enhancing the detection of small ligand of enzymatic reaction in BLM by electrochemical SPR spectroscopy.
本文首次运用电化学表面等离子体共振（SPR）方法，基于将辣根过氧化物酶（HRP）固定于由氧化还原聚苯胺（PAn）膜支撑的双层脂质膜（BLM）中来检测酶促反应。通过SPR动力学曲线原位监测HRP与PAn反应导致的PAn膜的氧化还原转变，利用电化学SPR光谱成功分析了HRP与H₂O₂的酶促反应。结果表明，这种支撑在PAn膜上的BLM不仅能保持固定在膜中的HRP的生物活性，还能为HRP与电极表面的PAn之间的电子转移提供通道。这些特性使得能够开发用于灵敏检测H₂O₂的SPR生物传感器。已通过电化学SPR光谱在5×10⁻⁵M至2×10⁻³M的浓度范围内检测到H₂O₂。每次检测后，通过将氧化的PAn电化学还原至其还原态，SPR传感器表面可完全再生。该方法为通过电化学SPR光谱增强对BLM中酶促反应的小配体的检测提供了一条新途径。
