采用纳米金作为抗体的固定基质,同时以硫堇为电活性指示剂,构建了一种免标记的电化学免疫传感器用于微囊藻毒素-LR(MC-LR)的灵敏检测。将萘酚修饰到金电极表面,通过静电作用将硫堇捕获。加入纳米金与硫堇的氨基结合,抗体将通过与纳米金作用而固定到电极表面。通过MC-LR与其相应抗体的特异性结合作用阻碍硫堇的电子传递,实现MC-LR的电化学检测。在最优实验条件下,MC-LR的质量浓度与电信号在5~100μg/L范围内呈良好的线性关系,检出限为0.71μg/L,可满足饮用水检测需求,能够用于实际水样中M C-LR的测定。
In this paper,by the use of gold nanoparticle as immobilization substrate for antibody and thionine( Thi) as the electro-active indicator,a label free electrochemical immunosensor has been fabricated for sensitive detection of microcystin-LR( MC-LR). Firstly,nafion was modified onto the gold electrode surface. Then,Thi could be captured through the electrostatic interaction. Then,gold nanoparticles were added to combine with the amino group of Thi. The antibody was immobilized onto the electrode surface through interaction with gold nanoparticle. Through the specific interaction between MC-LR and the corresponding antibody to inhibit the electron transfer of Thi,the electrochemical detection of MC-LR was achieved. Under the optimal experimental conditions,the concentration of MC-LR showed a good linear relationship against the electrochemical signal in the range from 5 ~ 100 μg/L. The limit of detection of MC-LR was 0. 71 μg/L,which met the required detection sensitivity of drinking water. At the same time, the experimental results indicated that the fabricated immunosensor could be used for the detection of MC-LR in real water samples. This electrochemical immunosensor is simple,without the need of complicate labeling procedure,and possesses good stability and reproducibility,thus has great importance for the monitor of MC-LR in the environment water samples.