Electrochemiluminescence biosensing of gene-specific methylation through magnetic capture and functional [Ru(byp)3]2+-doped silica
Electrochemiluminescence biosensing of gene-specific methylation through magnetic capture and functional [Ru(byp)3]2+-doped silica
复制标题
通过磁捕获和功能性 [Ru(byp)3]2 掺杂二氧化硅进行基因特异性甲基化的电化学发光生物传感
DOI：
10.1016/j.snb.2022.132857
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发表时间：
2022-10-21
影响因子：
8.4
通讯作者：
Ju，Huangxian
中科院分区：
文献类型：
--
作者：
Wu，Wenxin;Wu，Jie;Ju，Huangxian
本工作提出了一种电化学发光（ECL）生物传感策略，用于灵敏检测5-甲基胞嘧啶（5mC） DNA甲基化。将[Ru(byp)3]2+包封于二氧化硅纳米颗粒（Ru@SiO2）中，标记二抗（Ab2），制备信号探针，提供3.1 × 103[Ru(byp)3]2+进行信号放大。将生物素化的DNA固定在链霉亲和素修饰的磁性微珠（s - mb）上，以捕获顺序甲基化的ssDNA、5mC抗体和Ru@SiO2labeled Ab2。s - mb能够从复杂样品中的其他物种中快速分离捕获产物。以三丙胺为反应物，用Ru@SiO2的ECL信号读出甲基化DNA的浓度。该生物传感器对RASSF1A启动子区甲基化DNA序列的动态检测范围为0.5 ~ 50000 pM，检测限为0.1 pM。该方法具有灵敏度高、操作方便、易于自动化、通用性强等优点，无需核酸扩增和样品预处理即可检测重要基因特异性甲基化，潜力巨大。
This work proposed an electrochemiluminescence (ECL) biosensing strategy for sensitive detection of 5-methylcytosine (5mC) DNA methylation. The signaling probe was prepared by encapsulating [Ru(byp)3]2+in silica nanoparticle (Ru@SiO2) to label the secondary antibody (Ab2), which provided 3.1 × 103[Ru(byp)3]2+for signal amplification. Biotinylated DNA was immobilized on streptavidin-modified magnetic microbeads (S-MBs) to capture sequentially methylated ssDNA, 5mC antibody and Ru@SiO2labeled Ab2. S-MBs enable the rapid separation of the capture product from other species in complex samples. With tripropylamine as coreactant, the concentration of methylated DNA was read out with ECL signal of Ru@SiO2. The developed biosensor showed the dynamic range of 0.5–50,000 pM and a limit of detection of 0.1 pM for the methylated DNA sequence of the promoter region of RASSF1A. This proposed method possessed the advantages of high sensitivity, convenient operation, easy automation and excellent versatility, and enormous potential for the detection of important gene-specific methylation without nucleic acid amplification and sample pretreatment.