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高灵敏鲁米诺固定化方法及基于指数链取代反应的电化学发光策略用于监测细胞中 microRNA 的表达。

Highly-efficient luminol immobilization approach and exponential strand displacement reaction based electrochemiluminescent strategy for monitoring microRNA expression in cell.

机构信息

Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, China.

Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, China.

出版信息

Biosens Bioelectron. 2019 May 1;132:62-67. doi: 10.1016/j.bios.2019.02.005. Epub 2019 Feb 23.

Abstract

This work used 3,4,9,10-perylenetetracarboxylic acid-luminol composite (PTCA-luminol) as signal tag with improved ECL signal and applied cruciform DNA structure mediated exponential strand displacement reaction (SDR) to construct an ultrasensitive electrochemiluminescence (ECL) biosensor for microRNA-21 (miRNA-21) detection. The novel luminol-based signal tags was synthesized utilizing the π-π stacking interaction between PTCA and luminol, realizing highly effective and stable immobilization of luminol and resulting in good stability and strong ECL response. Meanwhile, target miRNA-21 triggered disaggregation of cruciform DNA structure was used to mediate exponential SDR for target recycling amplification. Taking advantage of the novel luminol-based signal tag and exponential SDR, the proposed ECL biosensor achieved excellent sensitivity with wide linear range from 10 aM to 100 pM and detection limit was 2 aM. Moreover, this ECL biosensor was applied to estimate the expression level of miRNA-21 and pharmacodynamics of matrine in human breast cancer cells (MCF-7 cells). The proposed biosensor provided a new opportunity for the preparation of ECL nanomaterials and exhibited great application potential in other biomarkers detection, clinical application and pharmacodynamics evaluation.

摘要

本工作使用 3,4,9,10-苝四羧酸-鲁米诺复合(PTCA-鲁米诺)作为信号标签,提高了 ECL 信号,并应用十字形 DNA 结构介导的指数链置换反应(SDR)构建了用于 microRNA-21(miRNA-21)检测的超灵敏电化学发光(ECL)生物传感器。新型基于鲁米诺的信号标签是利用 PTCA 和鲁米诺之间的π-π 堆积相互作用合成的,实现了鲁米诺的高效稳定固定,从而产生良好的稳定性和强 ECL 响应。同时,目标 miRNA-21 触发十字形 DNA 结构的解聚集,用于介导目标循环放大的指数 SDR。利用新型基于鲁米诺的信号标签和指数 SDR,所提出的 ECL 生物传感器具有出色的灵敏度,线性范围从 10 aM 到 100 pM,检测限为 2 aM。此外,该 ECL 生物传感器用于估计 miRNA-21 的表达水平和苦参碱在人乳腺癌细胞(MCF-7 细胞)中的药效动力学。该生物传感器为 ECL 纳米材料的制备提供了新的机会,并在其他生物标志物检测、临床应用和药效动力学评估中显示出巨大的应用潜力。

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