A simple and sensitive label-free fluorescent approach for protein detection based on a Perylene probe and aptamer
文献类型: 外文期刊
第一作者: Lv, Zhenzhen
作者: Lv, Zhenzhen;Liu, Jinchuan;Bai, Wenhui;Yang, Shuming;Chen, Ailiang;Lv, Zhenzhen;Liu, Jinchuan;Bai, Wenhui;Yang, Shuming;Chen, Ailiang;Lv, Zhenzhen
作者机构:
关键词: Aptamer;Perylene tetracarboxylic acid diimide;Thrombin;Label-free;Fluorescence
期刊名称:BIOSENSORS & BIOELECTRONICS ( 影响因子:10.618; 五年影响因子:9.323 )
ISSN:
年卷期:
页码:
收录情况: SCI
摘要: Highly sensitive detection of proteins is of great importance for effective clinical diagnosis and biomedical research. However, so far most detection methods rely on antibody-based immunoassays and are usually laborious and time-consuming with poor sensitivity. Here, we developed a simple and ultra-sensitive method to detect a biomarker protein-thrombin by taking advantage of the fluorescent probe Perylene tetracarboxylic acid diimide (PTCDI) derivatives and thrombin aptamer. The water-soluble dye PTCDI shows strong fluorescence in buffer solution for the existence of free dye monomer, but becomes weak after aggregation through self-assembly on nucleic acid aptamer. In the presence of thrombin, it specifically binds to thrombin aptamer which causes the conformational transition between aptamer and PTCDI and results in a significant fluorescence recovery. The results showed that as low as 40 pM of thrombin could be detected by this method. The high sensitivity of the developed sensing system mainly attributes to the ultra-sensitivity of the fluorescence intensity changes of PTCDI. With the specificity of aptamer, the assay exhibited high selectivity for thrombin against three other proteins (bovine serum albumin, lysozyme, mouse IgG) and 1% diluted fetal bovine serum. The detection method might be extended to sensitive detection of a variety of proteins for its advantages of isothermal conditions required, simple and rapid without multiple separation and washing steps. (C) 2014 Elsevier B.V. All rights reserved.
分类号: Q
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