Barton Andrew C, Davis Frank, Higson Séamus P J
Cranfield Health, Cranfield University, Silsoe, Beds, MK45 4DT, UK.
Anal Chem. 2008 Dec 15;80(24):9411-6. doi: 10.1021/ac801394d.
This paper describes the development and characterization of a label-less immunosensor for neuron-specific enolase (NSE) and its interrogation using an ac impedance protocol. Commercial screen-printed carbon electrodes were used as the basis for the sensor. Poly(1,2-diaminobenzene) was electrodeposited onto the sensors--and this modified surface was then sonochemically ablated to form an array of micropores. A second electropolymerization step was then used to deposit conductive polyaniline within these pores to give a microarray of polyaniline protrusions with diameters of several mum. This array was then utilized as a substrate to immobilize a biotinylated antibody for NSE using a classical avidin-biotin approach. Electrodes containing the antibodies were exposed to solutions of NSE and interrogated using an ac impedance protocol. The real component of the impedance of the electrodes was found to increase with increasing concentration of antigen. Control samples containing a nonspecific IgG antibody were also studied and calibration curves obtained by subtraction of the responses for specific and nonspecific antibody-based sensors, thereby accounting for and eliminating the effects of nonspecific adsorption of NSE. A linear relationship between the concentration of NSE in buffer solutions from 0 to 50 pg mL(-1) and the impedimetric response was observed.
本文描述了一种用于神经元特异性烯醇化酶（NSE）的无标记免疫传感器的开发与表征，以及使用交流阻抗协议对其进行检测的过程。使用商业丝网印刷碳电极作为传感器的基础。将聚（1,2 - 二氨基苯）电沉积到传感器上，然后对该修饰表面进行超声化学烧蚀以形成微孔阵列。接着进行第二步电聚合，在这些孔内沉积导电聚苯胺，得到直径为几微米的聚苯胺突出物微阵列。然后使用经典的抗生物素蛋白 - 生物素方法，将该阵列用作固定NSE生物素化抗体的底物。将含有抗体的电极暴露于NSE溶液中，并使用交流阻抗协议进行检测。发现电极阻抗的实部随抗原浓度的增加而增加。还研究了含有非特异性IgG抗体的对照样品，并通过减去基于特异性和非特异性抗体的传感器的响应来获得校准曲线，从而解释并消除了NSE非特异性吸附的影响。观察到缓冲溶液中NSE浓度在0至50 pg mL(-1)之间与阻抗响应呈线性关系。
