State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
Nucleic Acids Res. 2010 Nov;38(20):e185. doi: 10.1093/nar/gkq728. Epub 2010 Aug 19.
In this contribution, an electrochemical aptameric sensing scheme for the sensitive detection of small molecules is proposed using adenosine as a target model. A ferrocene (Fc)-functionalized thiolated aptamer probe is adapted and immobilized onto an electrode surface. Introducing a recognition site for EcoRI into the aptamer sequence not only suppresses the peak current corresponding to blank sample but also provides a signal-on response mechanism. In the absence of adenosine, the aptamer can fold into a hairpin structure and form a cleavable double-stranded region. Fc is capable of being removed from electrode surface by treatment with endonuclease, and almost no peak current is observed. The adenosine/aptamer binding induces the conformational transition of designed aptamer, dissociating the cleavable double-stranded segment. Therefore, the integrated aptamer sequence is maintained when exposing to endonuclease, generating a peak current of Fc. Utilizing the present sensing scheme, adenosine even at a low concentration can give a detectable current signal. Thus, a detection limit of 10(-10) M and a linear response range from 3.74×10(-9) to 3.74×10(-5) M are achieved. The proposed proof-of-principle of a novel electrochemical sensing is expected to extend to establish various aptameric platforms for the analysis of a broad range of target molecules of interest.
在本研究中,提出了一种基于电化学适体传感策略,用于灵敏检测小分子,以腺苷作为目标模型。适配并固定巯基化的适体探针于电极表面,该适体探针的一端连接有二茂铁(Fc)。在适体序列中引入 EcoRI 的识别位点,不仅抑制了空白样品对应的峰电流,而且提供了信号开启响应机制。在没有腺苷存在的情况下,适体可以折叠成发夹结构,并形成可切割的双链区域。Fc 可以通过内切酶处理从电极表面去除,几乎观察不到峰电流。腺苷/适体的结合诱导设计适体的构象转变,解离可切割的双链片段。因此,当暴露于内切酶时,整合的适体序列得以保持,产生 Fc 的峰电流。利用本传感策略,即使在低浓度下,腺苷也能给出可检测的电流信号。因此,检测限达到 10(-10) M,线性响应范围从 3.74×10(-9)到 3.74×10(-5) M。预期该新型电化学传感的原理验证将扩展到建立各种适体平台,用于分析广泛的感兴趣的目标分子。