Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, 221004 Xuzhou, China; Department of Neurology, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, China.
Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, 221004 Xuzhou, China.
Biosens Bioelectron. 2018 Jan 15;99:201-208. doi: 10.1016/j.bios.2017.07.063. Epub 2017 Jul 26.
In this paper, we reported a sensitive and selective electrochemical method for quantify DNA methylation, analyzing DNA MTase activity and screening of MTase inhibitor based on silver nanoparticles (Ag NPs) decorated carbon nanocubes (CNCs) as signal tag. The Ag NPs/CNCs was prepared by in situ growth of nanosilver on carboxylated CNCs and used as a tracing tag to label antibody. The sensor was prepared by immobilizing the double DNA helix structure on the surface of gold electrode. When DNA MTase was introduced, the probe was methylated. Successively, anti-5-methylcytosine antibody labeled Ag NPs/CNCs was specifically conjugated on the CpG methylation site. The electrochemical stripping signal of the Ag NPs was used to monitor the activity of MTase. The electrochemical signal has a linear relationship with M.SssI activities ranging from 0.05 to 120U/mL with a detection limit of 0.03U/mL. In addition, we also demonstrated the method could be used for rapid evaluation and screening of the inhibitors of MTase. The newly designed strategy avoid the requirement of deoxygenation for electrochemical assay, and thus provide a promising potential in clinical application.
在本文中,我们报道了一种基于银纳米粒子(Ag NPs)修饰的碳纳米立方体(CNCs)作为信号标记物,用于定量 DNA 甲基化、分析 DNA MTase 活性和筛选 MTase 抑制剂的灵敏选择性电化学方法。Ag NPs/CNCs 通过纳米银原位生长在羧基化的 CNCs 上制备,并用作标记抗体的示踪标记物。传感器通过将双链 DNA 结构固定在金电极表面来制备。当引入 DNA MTase 时,探针被甲基化。随后,特异性地将抗 5-甲基胞嘧啶抗体标记的 Ag NPs/CNCs 结合到 CpG 甲基化位点上。Ag NPs 的电化学剥离信号用于监测 MTase 的活性。电化学信号与 M.SssI 活性在 0.05 至 120U/mL 范围内呈线性关系,检测限为 0.03U/mL。此外,我们还证明了该方法可用于快速评估和筛选 MTase 的抑制剂。新设计的策略避免了电化学测定对脱氧的要求,因此在临床应用中具有广阔的应用前景。