Colorimetric determination of thrombin by exploiting a triple enzyme-mimetic activity and dual-aptamer strategy
Colorimetric determination of thrombin by exploiting a triple enzyme-mimetic activity and dual-aptamer strategy
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利用三重酶模拟活性和双适体策略比色测定凝血酶
DOI：
10.1007/s00604-017-2327-8
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发表时间：
2017-05
影响因子：
5.7
通讯作者：
Li Ding
中科院分区：
文献类型：
--
作者：
Lei Wang;Wen Yang;Tengfei Li;Duo Li;Zhimin Cui;Yan Wang;Shunli Ji;Qinxin Song;Chang Shu;Li Ding
**摘要**：本文描述了一种用于测定凝血酶的比色分析法。该方法基于三重模拟酶活性及双适配体结合策略。三重信号放大依赖于对显色底物的氧化……（原文“chromo”后似乎缺失内容，完整翻译需补充完整信息）
The article describes a colorimetric assay for the determination of thrombin. It is based on the application of a triple enzyme-mimetic activity and a dual aptamer binding strategy. The triple signal amplification relies on oxidation of the chromogenic enzyme substrate 3,3,5,5-tetramethylbenzidine (TMB) that is catalyzed by composites consisting of graphene oxide (GO), gold/platinum nanoparticles (AuPtNP), and aptamer (Apt15), a G-quadruplex/hemin conjugate. The dual-aptamer target binding strategy is based on the fact that thrombin has two active sites to be recognized by its aptamers (Apt15 and Apt29). Magnetic beads (MBs) were modified with Apt29 (Apt29-MB) and then are bound by the GO-AuPtNP-Apt15/G-quadruplex/hemin composites. In the presence of thrombin, Apt29-MB and the GO-AuPtNP-Apt15/G-quadruplex/hemin composites form a sandwich-like superstructure. Thus, the absorbance increases due to the formation of TMB oxide produced by catalysis of the composites. Under optimized conditions, the absorbance at 450 nm increases linearly in the 0.30 to 100 nM thrombin concentration range, and the limit of detection is 0.15 nM. The method is simple, rapid, and does not require complicated instrumentation. Bovine serum albumin, human serum albumin and other proteins were found not to interfere.Graphical abstractSchematic presentation of the photometric thrombin assay based on a triple enzyme-mimetic activity of combined nanomaterials (consisting of GO, AuPtNPs and the G-quadruplex/hemin DNAzyme) and two aptamers TMB: 3,3,5,5-tetramethylbenzidine, TMBox: 3,3,5,5-tetramethylbenzidine oxide, AuPtNP: gold/platinum nanoparticles).
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