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用于小分子高灵敏度信号开启检测的空白峰电流抑制电化学适体传感平台。

Blank peak current-suppressed electrochemical aptameric sensing platform for highly sensitive signal-on detection of small molecule.

机构信息

State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.

出版信息

Nucleic Acids Res. 2010 Nov;38(20):e185. doi: 10.1093/nar/gkq728. Epub 2010 Aug 19.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b231/2978380/3170b9ce40ce/gkq728s1.jpg

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