A solid-state electrochemiluminescence biosensing switch for detection of thrombin based on ferrocene-labeled molecular beacon aptamer

被引:108
作者
Wang, Xiaoying [1 ]
Dong, Ping [2 ]
Yun, Wen [2 ]
Xu, Ying [2 ]
He, Pingang [2 ]
Fang, Yuzhi [2 ]
机构
[1] Southeast Univ, Sch Publ Hlth, Nanjing 210009, Peoples R China
[2] E China Normal Univ, Dept Chem, Shanghai 200062, Peoples R China
关键词
Solid-state electrochemiluminescence biosensing switch; Molecular beacon aptamer; Thrombin; Ferrocene; Quenching effect; SENSITIVE DNA DETECTION; ELECTROGENERATED CHEMILUMINESCENCE; ELECTROCHEMICAL DETECTION; COMPOSITE FILMS; RU(BPY)(3)(2+); HYBRIDIZATION; PROTEIN; PROBES; GOLD; NANOPARTICLES;
D O I
10.1016/j.bios.2009.04.019
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A solid-state electrochemiluminescence (ECL) biosensing switch system based on special ferrocene-labeled molecular beacon aptamer (Fc-MBA) has been developed successfully for thrombin detections. Such special switch system includes two main parts, an ECL substrate and an ECL intensity switch. The ECL substrate was made by modifying the complex of Au nanoparticle and Ruthenium (II) tris-(bipyridine) (Ru(bpy)(3)(2+)-AuNPs) onto Au electrode. A molecular beacon aptamer labeled by ferrocene acted as the ECL intensity switch. The loop bases of the ECL intensity switch are designed with special anti-thrombin aptamer sequence which could be combined with its target protein via the reaction between aptamer and thrombin. During the reactions, the molecular beacon aptamer opened its stem-loop, and the labeled Fc was consequently kept away from the ECL substrate. Such structural change resulted in an obvious ECL intensity increment due to the decreased quenching effect of Fc to the ECL substrate. The analytic results are sensitive and specific. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:3288 / 3292
页数:5
相关论文
共 45 条
[31]   Luminescent supramolecular microstructures containing Ru(bpy)32+:: Solution-based self-assembly preparation and solid-state electrochemiluminescence detection application [J].
Sun, Xuping ;
Du, Yan ;
Zhang, Lixue ;
Dong, Shaojun ;
Wang, Erkang .
ANALYTICAL CHEMISTRY, 2007, 79 (06) :2588-2592
[32]   Molecular beacons: Probes that fluoresce upon hybridization [J].
Tyagi, S ;
Kramer, FR .
NATURE BIOTECHNOLOGY, 1996, 14 (03) :303-308
[33]   Multicolor molecular beacons for allele discrimination [J].
Tyagi, S ;
Bratu, DP ;
Kramer, FR .
NATURE BIOTECHNOLOGY, 1998, 16 (01) :49-53
[34]   Approaching real-time molecular diagnostics: Single-pair fluorescence resonance energy transfer (spFRET) detection for the analysis of low abundant point mutations in K-ras oncogenes [J].
Wabuyele, MB ;
Farquar, H ;
Stryjewski, W ;
Hammer, RP ;
Soper, SA ;
Cheng, YW ;
Barany, F .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (23) :6937-6945
[35]   Electrogenerated chemiluminescence detection for deoxyribonucleic acid hybridization based on gold nanoparticles carrying multiple probes [J].
Wang, Hui ;
Zhang, Chengxiao ;
Li, Yan ;
Qi, Honglan .
ANALYTICA CHIMICA ACTA, 2006, 575 (02) :205-211
[36]   A controllable solid-state Ru(bpy)32+ electrochemiluminescence film based on conformation change of ferrocene-labeled DNA molecular beacon [J].
Wang, Xiaoying ;
Yun, Wen ;
Dong, Ping ;
Zhou, Jingming ;
He, Pingang ;
Fang, Yuzhi .
LANGMUIR, 2008, 24 (05) :2200-2205
[37]   Solid-state electrochemiluminescence of tris(2,2′-bipyridyl) ruthenium [J].
Wei, Hui ;
Wang, Erkang .
TRAC-TRENDS IN ANALYTICAL CHEMISTRY, 2008, 27 (05) :447-459
[38]   Label free electrochemiluminescence protocol for sensitive DNA detection with a tris(2,2'-bipyridyl)ruthenium(II) modified electrode based on nucleic acid oxidation [J].
Wei, Hui ;
Du, Yan ;
Kang, Jianzhen ;
Wang, Erkang .
ELECTROCHEMISTRY COMMUNICATIONS, 2007, 9 (07) :1474-1479
[39]   ELECTROGENERATED CHEMI-LUMINESCENCE .41. ELECTROGENERATED CHEMI-LUMINESCENCE AND CHEMI-LUMINESCENCE OF THE RU(2,2'-BPY)3(2+)-S2O82- SYSTEM IN ACETONITRILE WATER SOLUTIONS [J].
WHITE, HS ;
BARD, AJ .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1982, 104 (25) :6891-6895
[40]   Molecular beacon aptamer fluoresces in the presence of Tat protein of HIV-1 [J].
Yamamoto, R ;
Kumar, PKR .
GENES TO CELLS, 2000, 5 (05) :389-396