Department of Chemistry, East China Normal University, Shanghai 200062, China.
Chemistry. 2010 Dec 27;16(48):14439-46. doi: 10.1002/chem.201001654.
α-Synuclein (α-SYN) is a very important neuronal protein that is associated with Parkinson's disease. In this paper, we utilized Au-doped TiO(2) nanotube arrays to design a photoelectrochemical immunosensor for the detection of α-SYN. The highly ordered TiO(2) nanotubes were fabricated by using an electrochemical anodization technique on pure Ti foil. After that, a photoelectrochemical deposition method was exploited to modify the resulting nanotubes with Au nanoparticles, which have been demonstrated to facilitate the improvement of photocurrent responses. Moreover, the Au-doped TiO(2) nanotubes formed effective antibody immobilization arrays and immobilized primary antibodies (Ab(1)) with high stability and bioactivity to bind target α-SYN. The enhanced sensitivity was obtained by using {Ab(2)-Au-GOx} bioconjugates, which featured secondary antibody (Ab(2)) and glucose oxidase (GOx) labels linked to Au nanoparticles for signal amplification. The GOx enzyme immobilized on the prepared immunosensor could catalyze glucose in the detection solution to produce H(2)O(2), which acted as a sacrificial electron donor to scavenge the photogenerated holes in the valence band of TiO(2) nanotubes upon irradiation of the other side of the Ti foil and led to a prompt photocurrent. The photocurrents were proportional to the α-SYN concentrations, and the linear range of the developed immunosensor was from 50 pg mL(-1) to 100 ng mL(-1) with a detection limit of 34 pg mL(-1). The proposed method showed high sensitivity, stability, reproducibility, and could become a promising technique for protein detection.
α-突触核蛋白(α-SYN)是一种非常重要的神经元蛋白,与帕金森病有关。在本文中,我们利用 Au 掺杂 TiO(2) 纳米管阵列设计了一种用于检测 α-SYN 的光电化学免疫传感器。高度有序的 TiO(2) 纳米管是通过在纯 Ti 箔上使用电化学阳极氧化技术制造的。之后,利用光电化学沉积法在所得纳米管上修饰 Au 纳米颗粒,这已被证明有助于提高光电流响应。此外,Au 掺杂的 TiO(2) 纳米管形成了有效的抗体固定阵列,并固定了具有高稳定性和生物活性的初级抗体(Ab(1))以结合靶标 α-SYN。通过使用{Ab(2)-Au-GOx} 生物缀合物获得了增强的灵敏度,该生物缀合物具有与 Au 纳米颗粒连接的二级抗体(Ab(2))和葡萄糖氧化酶(GOx)标记,用于信号放大。固定在制备的免疫传感器上的 GOx 酶可以在检测溶液中催化葡萄糖产生 H(2)O(2),H(2)O(2) 作为牺牲电子供体,在 Ti 箔的另一侧照射时清除 TiO(2) 纳米管价带中的光生空穴,导致光电流迅速增加。光电流与 α-SYN 浓度成正比,所开发的免疫传感器的线性范围为 50 pg mL(-1) 至 100 ng mL(-1),检测限为 34 pg mL(-1)。该方法具有高灵敏度、稳定性、重现性,有望成为蛋白质检测的一种有前途的技术。