Du Yan, Li Bingling, Wang Fuan, Dong Shaojun
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, PR China.
Biosens Bioelectron. 2009 Mar 15;24(7):1979-83. doi: 10.1016/j.bios.2008.10.019. Epub 2008 Nov 5.
We report a sensitively amplified electrochemical aptasensor using adenosine triphosphate (ATP) as a model. ATP is a multifunctional nucleotide that is most important as a "molecular currency" of intracellular energy transfer. In the sensing process, duplexes consisting of partly complementary strand (PCS1), ATP aptamer (ABA) and another partly complementary strand (PCS2) were immobilized onto Au electrode through the 5'-HS on the PCS1. Meanwhile, PCS2 was grafted with the Au nanoparticles (AuNPs) to amplify the detection signals. In the absence of ATP, probe methylene blue (MB) bound to the DNA duplexes and also bound to guanine bases specifically to produce a strong differential pulse voltammetry (DPV) signal. But when ATP exists, the ABA-PCS2 or ABA-PCS1 part duplexes might be destroyed, which decreased the amount of MB on the electrode and led to obviously decreased DPV signal. This phenomenon can be used to detect ATP and get a very sensitive detection limit low to 0.1nM, and the detection range could extend up to 10(-7)M. Compared to the sensing platform without PCS2 grafted AuNPs, amplified function of this sensing system was also evidently proved. Therefore, such PCS1-ABA-PCS2/AuNPs sensing system could provide a promising signal-amplified model for aptamer-based small-molecules detection.
我们报道了一种以三磷酸腺苷(ATP)为模型的灵敏放大电化学适配体传感器。ATP是一种多功能核苷酸,作为细胞内能量转移的“分子货币”最为重要。在传感过程中,由部分互补链(PCS1)、ATP适配体(ABA)和另一条部分互补链(PCS2)组成的双链体通过PCS1上的5'-HS固定在金电极上。同时,PCS2接枝金纳米颗粒(AuNPs)以放大检测信号。在不存在ATP的情况下,探针亚甲基蓝(MB)与DNA双链体结合,并特异性地与鸟嘌呤碱基结合,产生强烈的差分脉冲伏安法(DPV)信号。但是当ATP存在时,ABA-PCS2或ABA-PCS1部分双链体可能被破坏,这减少了电极上MB的量,导致DPV信号明显降低。这种现象可用于检测ATP,获得低至0.1 nM的非常灵敏的检测限,检测范围可扩展至10^(-7) M。与未接枝PCS2 AuNPs的传感平台相比,该传感系统的放大功能也得到了明显证明。因此,这种PCS1-ABA-PCS2/AuNPs传感系统可为基于适配体的小分子检测提供一种有前景的信号放大模型。