Dual-color determination of protein via terminal protection of small-molecule-linked DNA and the enzymolysis of exonuclease III
Dual-color determination of protein via terminal protection of small-molecule-linked DNA and the enzymolysis of exonuclease III
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通过小分子连接 DNA 的末端保护和核酸外切酶 III 的酶解双色测定蛋白质
DOI：
10.1016/j.bios.2014.02.060
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发表时间：
2014-08-15
影响因子：
12.6
通讯作者：
He, Zhike
中科院分区：
文献类型：
--
作者：
Chen, Chaohui;Xiang, Xia;He, Zhike
基于小分子连接DNA的末端保护和核酸外切酶III（ExoIII）的保护作用，我们开发了一种新型的双色荧光蛋白生物传感器。利用量子点发出的绿色荧光信号和[Ru（phen）（2）（dppx）]（2+）发出的红色荧光信号实现了对链霉亲和素（SA）的测定。在不存在SA的情况下，生物素-DNA被Exo III降解，从而使[Ru（phen）（2）（dppx）]（2+）用作量子点的荧光猝灭剂。SA的加入使生物素-双链DNA发生特异性结合，保护双链DNA与[Ru（phen）（2）（dppx）]（2+）结合，使量子点回收并产生[Ru（phen）（2）（dppx）]（2+）荧光。该传感器具有较高的灵敏度和较低的检测限（2.11 ng/mL），首次将双色量子点钌配合物用于蛋白质的检测。(c)2014爱思唯尔有限公司版权所有。
We have developed a new dual-color fluorescent biosensor for protein detection based on terminal protection of small-molecule-linked DNA and the enzymolysis of exonuclease III (Exo III). The determination of streptavidin (SA) was realized via fluorescence signals of the green color from quantum dots (QDs) and the red from [Ru(phen)(2)(dppx)](2+). In the absence of SA, biotin-DNA was degradated by the Exo III, thus making the [Ru(phen)(2)(dppx)](2+) employed as a fluorescence quencher to the QDs. With the addition of SA, dual-color response appeared because of the specific binding between SA and biotin so that the biotin-dsDNA was protected and combined with [Ru(phen)(2)(dppx)](2+), leading to the QDs recovery and the generating of [Ru(phen)(2)(dppx)](2+) fluorescence. This sensor exhibited high sensitivity with a low detection limit (2.11 ng/mL) and firstly introduced dual-color QDs ruthenium complex dyads to protein assay. (c) 2014 Elsevier B.V. All rights reserved.