Wang Xiaoying, Dong Ping, Yun Wen, Xu Ying, He Pingang, Fang Yuzhi
School of Public Health, Southeast University, Nanjing 210009, China.
Biosens Bioelectron. 2009 Jul 15;24(11):3288-92. doi: 10.1016/j.bios.2009.04.019. Epub 2009 Apr 22.
A solid-state electrochemiluminescence (ECL) biosensing switch system based on special ferrocene-labeled molecular beacon aptamer (Fc-MBA) has been developed successfully for thrombin detections. Such special switch system includes two main parts, an ECL substrate and an ECL intensity switch. The ECL substrate was made by modifying the complex of Au nanoparticle and Ruthenium (II) tris-(bipyridine) (Ru(bpy)(3)(2+)-AuNPs) onto Au electrode. A molecular beacon aptamer labeled by ferrocene acted as the ECL intensity switch. The loop bases of the ECL intensity switch are designed with special anti-thrombin aptamer sequence which could be combined with its target protein via the reaction between aptamer and thrombin. During the reactions, the molecular beacon aptamer opened its stem-loop, and the labeled Fc was consequently kept away from the ECL substrate. Such structural change resulted in an obvious ECL intensity increment due to the decreased quenching effect of Fc to the ECL substrate. The analytic results are sensitive and specific.
一种基于特殊二茂铁标记分子信标适配体(Fc-MBA)的固态电化学发光(ECL)生物传感开关系统已成功开发用于凝血酶检测。这种特殊的开关系统包括两个主要部分,一个ECL底物和一个ECL强度开关。ECL底物是通过将金纳米颗粒与三(联吡啶)钌(II)(Ru(bpy)(3)(2+)-AuNPs)的复合物修饰到金电极上制成的。由二茂铁标记的分子信标适配体充当ECL强度开关。ECL强度开关的环碱基设计有特殊的抗凝血酶适配体序列,该序列可通过适配体与凝血酶之间的反应与其靶蛋白结合。在反应过程中,分子信标适配体打开其茎环结构,标记的Fc因此远离ECL底物。由于Fc对ECL底物的猝灭作用降低,这种结构变化导致ECL强度明显增加。分析结果灵敏且特异。