Tannic acid-accelerated fenton chemical reaction amplification for fluorescent biosensing: The proof-of-concept towards ultrasensitive detection of DNA methylation

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL

Talanta Pub Date : 2023-06-12 DOI:10.1016/j.talanta.2023.124811

Hongding Zhang , Sifei Wu , Zhixiao Song , Linxia Fang , Hai-Bo Wang

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引用次数: 0

Abstract

As a promising biomarker, the level of methylated DNA usually changes in the early stage of the cancer. Ultrasensitive detection of the changes of methylated DNA offers possibility for early diagnosis of cancer. In this work, a tannic acid-accelerated Fenton chemical reaction amplification was firstly proposed for the construction of ultrasensitive fluorescent assay. Tannic acid was used as reductant to accelerate Fenton reaction procedure through the conversion of Fe³⁺/Fe²⁺, generating hydroxyl radicals (·OH) continuously. The produced ·OH oxidized massive non-fluorescent terephthalic acid (TA) to fluorescent-emitting hydroxy terephthalic acid (TAOH). In this way, the fluorescent signal could be greatly enhanced and the sensitivity was improved almost 116 times. The proposed signal amplification strategy was further applied to detect of DNA methylation with the assistance of liposome encapsulated with tannic-Fe³⁺ complexes. The methylated DNA was firstly captured through the hybridization with its complementary DNA that were pre-modified in the 96-well plate via the combination between streptavidin (SA) and biotin. Then, 5 mC antibody on the surface of liposomes specially recognized and combined with methylation sites, which brought large amount of tannic-Fe³⁺ complexes to participate Fenton reaction. The fluorescence of generated TAOH was depended on the concentration of methylated DNA. The assay showed good analytical performance for methylated DNA with a limit of detection (LOD) of 1.4 fM. It's believed that tannic acid-accelerated Fenton chemical reaction amplification strategy provides a promising platform for ultrasensitive fluorescent detection of low abundant biomarkers.

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单宁酸加速芬顿化学反应扩增用于荧光生物传感:对DNA甲基化超灵敏检测的概念证明

作为一种很有前景的生物标志物，甲基化DNA的水平通常在癌症的早期发生变化。甲基化DNA变化的超灵敏检测为癌症的早期诊断提供了可能。本文首次提出了单宁酸加速芬顿化学反应扩增技术，用于构建超灵敏荧光检测。以单宁酸为还原剂，通过Fe3+/Fe2+的转化加速Fenton反应过程，不断生成羟基自由基(·OH)。生成的·OH将大量非荧光对苯二甲酸(TA)氧化为荧光发射的羟基对苯二甲酸(TAOH)。这样可以大大增强荧光信号，灵敏度提高近116倍。在单宁- fe3 +配合物包裹的脂质体的帮助下，进一步应用所提出的信号放大策略检测DNA甲基化。甲基化DNA首先通过与互补DNA的杂交捕获，互补DNA通过链亲和素(SA)和生物素的结合在96孔板上预先修饰。然后脂质体表面的5mc抗体特异性识别并结合甲基化位点，带来大量单宁- fe3 +配合物参与Fenton反应。生成的TAOH的荧光与甲基化DNA的浓度有关。该方法对甲基化DNA具有良好的分析性能，检测限(LOD)为1.4 fM。认为单宁酸加速芬顿化学反应扩增策略为低丰度生物标志物的超灵敏荧光检测提供了一个有前景的平台。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.