Time-resolved fluorescence aptamer-based sandwich assay for thrombin detection

被引:48
作者
Huang, Da-Wei [1 ]
Niu, Cheng-Gang [1 ,2 ]
Qin, Pin-Zhu [1 ]
Ruan, Min [1 ]
Zeng, Guang-Min [1 ]
机构
[1] Hunan Univ, Coll Environm Sci & Engn, Key Lab Environm Biol & Pollut Control, Minist Educ, Changsha 410082, Hunan, Peoples R China
[2] Cent S Univ, State Key Lab Powder Met, Changsha 410083, Peoples R China
基金
高等学校博士学科点专项科研基金; 中国国家自然科学基金;
关键词
Thrombin detection; Aptamer; Time resolved fluorescence; Eu(TTA)3(5-NH2-phen); IN-VITRO SELECTION; QUANTUM-DOT; BINDING; MOLECULES; NANOPARTICLES; LIGANDS;
D O I
10.1016/j.talanta.2010.09.004
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
In the present study the authors report a novel sensitive method for the detection of thrombin using time-resolved fluorescence sensing platform based on two different thrombin aptamers The thrombin 15-mer aptamer as a capture probe was covalently attached to the surface of glass slide and the thrombin 29-mer aptamer was fluorescently labeled as a detection probe A bifunctional europium complex was used as the fluorescent label The introduction of thrombin triggers the two different thrombin aptamers and thrombin to form a sandwich structure The fluorescence intensity is proportional to the thrombin concentration The present sensing system could provide both a wide linear dynamic range and a low detection limit The proposed sensing system also presented satisfactory specificity and selectivity Results showed that thrombin was retained at the aptamer-modified glass surface while nonspecific proteins were removed by rinsing with buffer solution This approach successfully showed the suitability of aptamers as low molecular weight receptors on glass slides for sensitive and specific protein detection (C) 2010 Elsevier B V All rights reserved
引用
收藏
页码:185 / 189
页数:5
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