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基于苝修饰的银微花作为新型电化学发光指示剂构建用于生物传感应用的高效DNA放大器。

Engineering a high-efficient DNA amplifier for biosensing application based on perylene decorated Ag microflowers as novel electrochemiluminescence indicators.

作者信息

Yang Fang, He Yan-Wei, Chai Ya-Qin, Yuan Ruo, Zhuo Ying

机构信息

Key Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, PR China.

Key Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, PR China.

出版信息

Biosens Bioelectron. 2021 Jun 15;182:113178. doi: 10.1016/j.bios.2021.113178. Epub 2021 Mar 21.

Abstract

DNA-based amplifiers with high programmability and accurate molecular recognition ability have become a versatile platform for target amplification. However, the random diffusion of capture probes (CPs) in most DNA amplifiers limits the target recognition efficiency, affecting the limit of detection. Herein, a high-efficient DNA amplifier was developed by localizing the CPs consisted of the unique palindromic tails and target recognition sequences on Au nanoparticle modified magnetic beads (Au@MBs). In the presence of target K-ras gene, the CPs with high local concentration and orientation could capture the target efficiently to expose their palindromic tails, which could act as primers to trigger the polymerization for target recycling. More importantly, the polymerization products could involve in the next recycle and produce abundant mimic targets (MTs) continuously, thereby achieving the detection of trace K-ras gene. Meanwhile, a novel electrochemiluminescence (ECL) indicator of a thin-layer of perylene (Pe) molecules decorated Ag microflowers (Pe@Ag MFs) was obtained based on the reaction between the perylene cation radical (Pe) and Ag atoms. The obtained Pe@Ag MFs exhibited desirable ECL performance because (i) a thin-layer of Pe molecules could reduce the inner filter effect and inactive emitters, (ii) the Ag MFs as coreaction accelerator could react with SO to produce more SO and shorten the distance between Pe and SO to significantly enhance the ECL intensity of Pe with less energy loss. This work paves the way for the development of efficient amplification strategy and offers a paradigm for the preparation of high-efficiency ECL indicators.

摘要

具有高可编程性和精确分子识别能力的基于DNA的放大器已成为用于目标扩增的通用平台。然而,大多数DNA放大器中捕获探针(CPs)的随机扩散限制了目标识别效率,影响了检测限。在此,通过将由独特回文尾和目标识别序列组成的CPs定位在金纳米颗粒修饰的磁珠(Au@MBs)上,开发了一种高效DNA放大器。在目标K-ras基因存在的情况下,具有高局部浓度和取向的CPs可以有效地捕获目标,从而暴露其回文尾,这些回文尾可以作为引物触发聚合反应以实现目标循环利用。更重要的是,聚合产物可以参与下一次循环并持续产生大量模拟目标(MTs),从而实现对痕量K-ras基因的检测。同时,基于苝阳离子自由基(Pe)与银原子之间的反应,获得了一种新型的电化学发光(ECL)指示剂,即由苝(Pe)分子修饰的银微花(Pe@Ag MFs)薄层。所制备的Pe@Ag MFs表现出理想的ECL性能,原因如下:(i)苝分子薄层可以降低内滤效应和非活性发射体;(ii)作为共反应促进剂的银微花可以与SO反应生成更多的SO,并缩短Pe与SO之间的距离,从而以较少的能量损失显著增强Pe的ECL强度。这项工作为高效扩增策略的发展铺平了道路,并为制备高效ECL指示剂提供了范例。

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