Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Hubei Province Engineering and Technology Research Center for Fluorinated Pharmaceuticals, and Wuhan University School of Pharmaceutical Sciences, Wuhan 430071, China.
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Hubei Province Engineering and Technology Research Center for Fluorinated Pharmaceuticals, and Wuhan University School of Pharmaceutical Sciences, Wuhan 430071, China.
Bioelectrochemistry. 2022 Aug;146:108111. doi: 10.1016/j.bioelechem.2022.108111. Epub 2022 Mar 28.
The detection of methylation level in O-methylguanine-DNA methyltransferase (MGMT) gene is of great significance for diagnosis, prognosis and treatment of glioblastoma. Herein, we proposed an electrochemical immunosensor for detection of MGMT gene methylation. Graphene oxide-magnetic nanoparticles-β-cyclodextrin (GO-FeO-β-CD) nanocomposite was covered on electrode, and then anti-5-methylcytosine (5mC) antibody was immobilized on modified electrode via host-guest interaction with β-CD. The immobilized antibody specifically recognized and directly captured 5mC on DNA containing MGMT promoter sequence (MGMT-DNA). The methylation level in MGMT-DNA could be evaluated using Ru(NH) as an electrochemical signal indicator. The redox recycling of Ru(NH) was triggered by Fe(CN) and further amplified current response. A detection limit as low as 0.0825 pM was obtained. Finally, this electrochemical method was successfully applied to analysis of methylated MGMT-DNA in biological sample and detection of other methylation modifications, such as N6-methyladenosine in RNA.
检测 O-甲基鸟嘌呤-DNA 甲基转移酶（MGMT）基因的甲基化水平对胶质母细胞瘤的诊断、预后和治疗具有重要意义。本文提出了一种用于检测 MGMT 基因甲基化的电化学免疫传感器。将氧化石墨烯-磁性纳米粒子-β-环糊精（GO-FeO-β-CD）纳米复合材料覆盖在电极上，然后通过 β-CD 的主体-客体相互作用将抗 5-甲基胞嘧啶（5mC）抗体固定在修饰电极上。固定化抗体特异性识别并直接捕获含有 MGMT 启动子序列（MGMT-DNA）的 DNA 上的 5mC。使用 Ru(NH)作为电化学信号指示剂来评估 MGMT-DNA 中的甲基化水平。Fe(CN)触发 Ru(NH)的氧化还原循环，并进一步放大电流响应。该方法的检测限低至 0.0825 pM。最后，该电化学方法成功应用于生物样品中甲基化 MGMT-DNA 的分析以及 RNA 中其他甲基化修饰（如 N6-甲基腺苷）的检测。
