脱氧核酶
血红素
G-四倍体
适体
化学
检出限
靶蛋白
组合化学
比色法
蛋白质检测
分子信标
DNA
生物化学
生物物理学
核酸
寡核苷酸
纳米技术
色谱法
分子生物学
酶
生物
基因
材料科学
血红素
作者
Hao Wu,Kai Zhang,Yaling Liu,Hongyong Wang,Jun Wu,Feifan Zhu,Zingway Pei
标识
DOI:10.1016/j.bios.2014.09.096
摘要
In this work, a new binding-induced and label-free colorimetric method for protein detection has been developed on the basis of an autonomous assembly of hemin/G-quadruplex DNAzyme amplification strategy. The system consists of two proximity probes carrying two aptamer sequences as recognition elements for target, and two hairpin structures include three-fourths and one-fourth of the G-quadruplex sequences in inactive configuration as functional elements. In the presence of target protein, two proximity probes bind to the protein simultaneously, forming a stable DNA–protein complex. Then the complex triggers an autonomous cross-opening of the two functional hairpin structures, leading to the formation of numerous hemin/G-quadruplex DNAzymes. The resulting DNAzymes catalyze the oxidation of colorless 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS2−) to the green-colored ABTS•− with the presence of H2O2, thus providing the amplified colorimetric detection of target. Using human α-thrombin as the protein target, this binding-induced DNAzyme amplification colorimetric method affords high sensitivity with a detection limit of 1.9 pM. Furthermore, this method might be further extended to sensitive detection of other proteins by simply replacing recognition elements of proximity probes.
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