采用电沉积方法将Pd纳米颗粒沉积到玻碳电极(GCE)表面,再将Pd纳米颗粒修饰电极插入H2SO4溶液中,吸收适量活性氢后,转移到HAuCl4溶液中,静置一定时间后,使金被活性氢还原并自发沉积到Pd纳米颗粒修饰的玻碳电极表面。通过自组装作用将带巯基的凝血酶适配体Ⅰ固定在Pd-Au/GCE表面,制得非标记型凝血酶适配体传感器。当凝血酶与凝血酶适配体结合时,覆盖在电极表面,从而阻碍了电极表面的Pd-Au纳米颗粒对H2O2的催化还原活性,通过监测H2O2还原电流的减小程度,实现对凝血酶的定量检测。考察了pH值、培育时间等实验条件对响应电流的影响以及Pd-Au纳米颗粒的协同作用。实验表明,此传感器的线性范围为3.0~300 nmol/L,检出限为0.98 nmol/L。
Pd nanoparticles (Pd NPs) were electrodeposited on the glassy carbon electrode (GCE). Then, Pd NPs/GCE was further inserted into H2 SO4 solution to polarize for 5 min to absorb a certain amount of active hydrogen. Then, the electrode was quickly inserted to HAuCl4 solution for 15 min. AuNPs were deposited spontaneously on the surface of Pd NPs due to the reduction of active hydrogen. As a result, Pd-AuNPs were modified on the surface of GCE. Next, aptamer I of thrombin was immobilized on Pd-Au nanoparticles. In the presence of thrombin, it bound with the aptamer immobilized on Pd-Au nanoparticles and thus the formed complex obstructed the catalysis of Pd-Au nanoparticles to H2 O2 . Hence, the reduction current of H2 O2 decreased with the increase of thrombin concentration. The aptamer sensor had a good linear relationship with the concentration of thrombin in the range of 2. 98-297 nmol/L with a detection limit of 0. 98 nmol/L.