Wu Zai-Sheng, Guo Meng-Meng, Zhang Song-Bai, Chen Chen-Rui, Jiang Jian-Hui, Shen Guo-Li, Yu Ru-Qin
State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, P. R. China.
Anal Chem. 2007 Apr 1;79(7):2933-9. doi: 10.1021/ac0622936. Epub 2007 Mar 6.
Aptamers are nucleic acids that have high affinity and selectivity for their target molecules. A target may induce the structure switching from a DNA/DNA duplex to a DNA/target complex. In the present study, a reusable electrochemical sensing platform based on structure-switching signaling aptamers for highly sensitive detection of small molecules is developed using adenosine as a model analyte. A gold electrode is first modified with polytyramine and gold nanoparticles. Then, thiolated capture probe is assembled onto the modified electrode surface via sulfur-gold affinity. Ferrocene (Fc)-labeled aptamer probe, which is designed to hybridize with capture DNA sequence and specifically recognize adenosine, is immobilized on the electrode surface by hybridization reaction. The introduction of adenosine triggers structure switching of the aptamer. As a result, Fc-labeled aptamer probe is forced to dissociate from the sensing interface, resulting in a decrease in redox current. The decrement of peak current is proportional to the amount of adenosine. The present sensing system could provide both a wide linear dynamic range and a low detection limit. In addition, high selectivity, good reproducibility, stability, and reusability are achieved. The recovery test demonstrates the feasibility of the designed sensing system for an adenosine assay.
适配体是对其靶分子具有高亲和力和选择性的核酸。靶标可诱导结构从DNA/DNA双链体转变为DNA/靶标复合物。在本研究中,以腺苷作为模型分析物,开发了一种基于结构转换信号适配体的可重复使用的电化学传感平台,用于高灵敏度检测小分子。首先用聚酪胺和金纳米颗粒修饰金电极。然后,通过硫-金亲和力将硫醇化捕获探针组装到修饰电极表面。二茂铁(Fc)标记的适配体探针被设计用于与捕获DNA序列杂交并特异性识别腺苷,通过杂交反应固定在电极表面。腺苷的引入触发适配体的结构转换。结果,Fc标记的适配体探针被迫从传感界面解离,导致氧化还原电流降低。峰值电流的降低与腺苷的量成正比。本传感系统可提供宽线性动态范围和低检测限。此外,还实现了高选择性、良好的重现性、稳定性和可重复使用性。回收率测试证明了所设计的传感系统用于腺苷测定的可行性。