UMR-CNRS 8182, ICMMO, University Paris-Sud, Orsay, France.
Anal Chem. 2013 Aug 20;85(16):7704-12. doi: 10.1021/ac400605p. Epub 2013 Aug 1.
The present work aims to develop an electrochemical biosensor based on aptamer able to detect human cellular prions PrP(C) as a model biomarker of prion disease with high sensitivity. We designed the biosensor using multiwalled carbon nanotubes (MWCNTs) modified with polyamidoamine dendrimers of fourth generation (PAMAM G4) which in turn were coupled to DNA aptamers used as bioreceptors. Electrochemical signal was detected by a ferrocenyl redox marker incorporated between the dendrimers and aptamers interlayer. MWCNTs, thanks to their nanostructure organization and electrical properties, allow the distribution of aptamers and redox markers over the electrode surface. We demonstrated that the interaction between aptamers and prion proteins leads to variation in the electrochemical signal of the ferrocenyl group. High sensitivity with a detection limit of 0.5 pM and a wide linear range of detection from 1 pM to 10 μM has been demonstrated. Detection of PrP(C) in spiked blood plasma has been achieved in the same range of concentrations as for detection of PrP(C) in buffer. The sensor demonstrated a recovery of minimum 85% corresponding to 1 nM PrP(C) and a maximum of 127% corresponding to 1 pM PrP(C).
本工作旨在开发一种基于适配体的电化学生物传感器,能够以高灵敏度检测人类细胞朊病毒 PrP(C),作为朊病毒病的模型生物标志物。我们使用经第四代聚酰胺胺树状大分子 (PAMAM G4) 修饰的多壁碳纳米管 (MWCNTs) 设计了该生物传感器,而后者又与用作生物受体的 DNA 适配体偶联。电化学信号通过嵌入在树突状聚合物和适配体夹层之间的二茂铁氧化还原标记物来检测。MWCNTs 由于其纳米结构组织和电性能,允许适配体和氧化还原标记物在电极表面分布。我们证明了适配体与朊病毒蛋白之间的相互作用会导致二茂铁基团的电化学信号发生变化。该生物传感器具有高灵敏度,检测限为 0.5 pM,检测范围从 1 pM 到 10 μM 很宽。在与缓冲液中检测相同浓度范围内,已实现了对添加的血漿中的 PrP(C) 的检测。该传感器的回收率至少为 85%,对应于 1 nM PrP(C,最高为 127%,对应于 1 pM PrP(C)。