Labeless Immunosensor Assay for the Stroke Marker Protein Neuron Specific Enolase Based upon an Alternating Current Impedance Protocol
Labeless Immunosensor Assay for the Stroke Marker Protein Neuron Specific Enolase Based upon an Alternating Current Impedance Protocol
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DOI：
10.1021/ac801394d
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发表时间：
2008-12-15
影响因子：
7.4
通讯作者：
Higson, Seamus P. J.
中科院分区：
文献类型：
--
作者：
Barton, Andrew C.;Davis, Frank;Higson, Seamus P. J.
本文介绍了神经元特异性烯醇化酶(NSE)无标记免疫传感器的研制和表征，并用交流阻抗法对其进行了检测。该传感器以商用丝网印刷碳电极为基础。将聚(1，2-二氨基苯)电沉积到传感器上，然后对修饰后的表面进行超声化学消融，形成一系列微孔。然后使用第二电聚合步骤在这些孔内沉积导电聚苯胺，以得到直径为几个pm的聚苯胺突起的微阵列然后利用该阵列作为底物，使用经典的亲和素-生物素方法固定NSE的生物素化抗体。将含有抗体的电极暴露在NSE溶液中，并使用交流阻抗方案进行询问。发现电极阻抗的实际分量随着抗原浓度的增加而增加。含有非特异性抗体的对照样品也被研究，并通过减去特异性和非特异性抗体传感器的反应获得校准曲线，从而解释和消除NSE的非特异性吸附的影响。在0~50pgmL(-1)的缓冲溶液中，NSE的浓度与阻抗响应呈线性关系。
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 pm. 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.