Electrogenerated chemiluminescence biosensing method for the detection of DNA demethylase activity: Combining MoS2 nanocomposite with DNA supersandwich
Electrogenerated chemiluminescence biosensing method for the detection of DNA demethylase activity: Combining MoS2 nanocomposite with DNA supersandwich
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检测DNA去甲基酶活性的电化学发光生物传感方法：MoS2纳米复合材料与DNA超三明治的结合
DOI：
10.1016/j.snb.2017.01.065
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发表时间：
2017-06
期刊：
影响因子：
--
通讯作者：
Qi Honglan
中科院分区：
文献类型：
--
作者：
Sun Huiping;Ma Shangxian;Li Yan;Qi Honglan
本文报道了一种基于Hpa Ⅱ酶切结合硫堇-二硫化钼（Th-MoS 2）纳米复合物和超夹心DNA信号放大的超灵敏电致化学发光（ECL）生物传感方法。硫堇（Thionin，Th）作为一种有效的介体，将捕获探针固定在MoS 2表面，然后，含有甲基化位点5′-CmCGG-3′的长串联体与捕获探针杂交，形成超夹心DNA结构。Ru（phen）32+ECL试剂可以以高亲和力嵌入到所形成的超夹层DNA结构的凹槽中，导致增强的信号传导。在生物传感电极（Ru（phen）32+/supersandwich DNA/Th-MoS 2/玻碳电极）上，去甲基化后的supersandwich DNA能被HpaII特异性切割，从而降低Ru（phen）32+的电化学发光信号。对于DNA去甲基化酶活性的检测，达到了0.16 pg mL− 1的极低检测限。这项工作表明，使用二硫化钼-硫堇纳米复合物和supersandwich与固有的灵敏度ECL方法的双信号放大的组合是一个很有前途的策略，用于检测DNA去甲基化和测定去甲基化酶的活性。
We report an ultrasensitive electrogenerated chemiluminescence (ECL) biosening method for the detection of DNA demethylase activity based on the site-specific cleavage of theHpaII endonuclease in conjunction with signal amplification by a Thionin-MoS2(Th-MoS2) nanocomposite and supersandwich DNA. The Thionin (Th) served as an effective mediator for the immobilization of the capture probe onto the surface of MoS2, after which the long concatamers containing the methylated site 5′-CmCGG-3′ were hybridized with the capture probe, creating a supersandwich DNA structure. Ru(phen)32+ECL reagents can be intercalated into the grooves of the formed supersandwich DNA structure with high affinity leading to enhanced signaling. At the biosensing electrode (Ru(phen)32+/supersandwich DNA/Th-MoS2/glassy carbon electrode) after demethylation with demethylase, the demethylated supersandwich DNA could be specifically cleaved usingHpaII, resulting in a decreased ECL signal from Ru(phen)32+. An extremely low detection limit of 0.16 pg mL−1was achieved for the detection of DNA demethylase activity. This work demonstrates that the combination of dual signal amplification using a MoS2-thionin nanocomposite and supersandwich with an inherently sensitive ECL method is a promising tactic for the detection of DNA demethylation and assay of the activity of demethylase.
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影响因子：
12.6
作者：
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DOI：
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