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将抗体结合蛋白A和交联剂联硫基二(琥珀酰亚胺丙酸盐)(DTSP)组成的骨架修饰到电极表面,再将单克隆雌二醇抗体与骨架中的蛋白A相结合,制备出电化学免疫传感器.利用样品雌二醇与辣根过氧化酶所标记的雌二醇同传感器表面抗体的竞争性结合,将该传感器用于测定溶液中样品雌二醇的浓度;利用方波伏安法监测电化学还原辣根过氧化酶催化产生的苯醌分子的电流,以还原电流为纵坐标,以雌二醇浓度为横坐标绘制标准曲线.结果表明,雌二醇浓度在50～1 500ng.L-1范围内与方波伏安还原电流呈良好的线性关系,灵敏度为0.51μA.ng-1.L,检测限为50ng.L-1.所制备的免疫传感器良好的分析性能得益于蛋白A和DTSP所组成的骨架,该骨架能够增加免疫分子组分在电极表面的修饰量,并能够控制抗体在电极上的结合方位,使其抗原结合位点朝向电极外端,减少结合空间阻碍.
The skeleton composed of the antibody binding protein A and the cross-linking agent dithiobis (succinimidylpropionate) (DTSP) was modified to the electrode surface, and then the monoclonal estradiol antibody was bound to the protein A in the skeleton, Electrochemical immunosensor was prepared by using the sample estradiol and horseradish peroxidase labeled estradiol with the sensor surface antibody binding, the sensor was used to determine the concentration of the sample solution of estradiol; The wave voltammetry was used to monitor the electrochemical reduction of benzoquinone molecules catalyzed by horseradish peroxidase and the standard curve was drawn with the reduction of current as the ordinate and the concentration of estradiol as the abscissa.The results showed that the estradiol concentration was 50 ~ 1 500ng.L-1 and the square wave voltammetry reduction current showed a good linear relationship with a sensitivity of 0.51μA.ng-1.L, a detection limit of 50ng.L-1. The prepared immunosensor good analysis Performance thanks to the skeleton composed of protein A and DTSP, the skeleton can increase the amount of modification of the immune molecule components on the electrode surface, and can control the binding orientation of the antibody on the electrode, the antigen binding site toward the outer end of the electrode, Reduce the combination of space obstruction.