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稳定的硝酰基氮氧自由基单自由基 MATMP 作为新型可逆加成断裂链转移 (RAFT) 聚合单体用于超灵敏 DNA 检测。

Stable nitronyl nitroxide monoradical MATMP as novel monomer of reversible addition fragmentation chain transfer (RAFT) polymerization for ultrasensitive DNA detection.

机构信息

School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, Jiangsu, 210094, PR China.

School of Environmental Science, Nanjing Xiaozhuang University, Nanjing, 211171, PR China.

出版信息

Anal Chim Acta. 2022 Aug 22;1222:340167. doi: 10.1016/j.aca.2022.340167. Epub 2022 Jul 14.

Abstract

In this work, it came up a hydrophilic and stable nitronyl nitroxide monoradical 4-methacryloyloxy-2,2,6,6-tetramethylpiperidine 1-oxyl freeradical (MATMP) as new monomer of polymerization for DNA detection. The detection limit was over 1,000,000 times lower than 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) derivatives as electrochemical labels alone. Within this approach, a single biomolecule can be converted into the strong electrochemical signal, therefore lung cancer DNA can be detected at low concentration. For the first step, the HS-PNA probe was fixed on the surface of the Au electrode. After the target DNA was captured by complementary base pairing, 4-cyano-4-(phenylcarbonothioylthio) pentanoic acid (CPAD), chain transfer agent of RAFT polymerization, was bonded to target DNA as reaction via coordination bond of Zr. Electroactive polymers had grown by means of surface initiated thermally RAFT polymerization with MATMP as monomer. MATMP polymer significantly improves the electrochemical signal. This method can detect DNA from 0.01 fM to 10 pM, and detection limit is 1.51 aM. The sensitivity of this method is greater than that in most other reported signal amplification strategies of DNA biosensor, which indicates that it is appropriate for single nucleotide polymorphism analysis and will broaden prospects for biological molecules detection application.

摘要

在这项工作中,我们提出了一种亲水性和稳定性的硝酰氮自由基单自由基 4-丙烯酰氧基-2,2,6,6-四甲基哌啶 1-氧自由基(MATMP)作为聚合新单体用于 DNA 检测。检测限比单独的 2,2,6,6-四甲基哌啶-1-氧(TEMPO)衍生物高 100 多万倍。在这种方法中,单个生物分子可以转化为强电化学信号,因此可以在低浓度下检测肺癌 DNA。在第一步中,HS-PNA 探针被固定在 Au 电极的表面。当目标 DNA 通过互补碱基配对被捕获后,作为反应的链转移剂 4-氰基-4-(苯甲酰硫代)戊酸(CPAD)通过 Zr 的配位键与目标 DNA 结合。通过表面引发的热 RAFT 聚合,以 MATMP 为单体,生长了电活性聚合物。MATMP 聚合物显著提高了电化学信号。该方法可以从 0.01 fM 到 10 pM 检测 DNA,检测限为 1.51 aM。该方法的灵敏度大于大多数其他报道的 DNA 生物传感器信号放大策略的灵敏度,这表明它适用于单核苷酸多态性分析,并将拓宽生物分子检测应用的前景。

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