Bifunctional combined aptamer for simultaneous separation and detection of thrombin

被引:40
作者
Bing, Tao [1 ,2 ]
Liu, Xiangjun [1 ]
Cheng, Xiaohong [1 ,2 ]
Cao, Zehui [1 ]
Shangguan, Dihua [1 ]
机构
[1] Chinese Acad Sci, Beijing Natl Lab Mol Sci, Inst Chem, Beijing 100190, Peoples R China
[2] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
关键词
Aptamer; Biosensor; Protein detection; Separation; Thrombin; Streptavidin; STRANDED-DNA MOLECULES; FLUORESCENCE ANISOTROPY; HIGH-AFFINITY; RNA APTAMERS; SELECTION; BIND; LIGANDS; SENSORS; D(GGTTGGTGTGGTTGG); OLIGONUCLEOTIDES;
D O I
10.1016/j.bios.2009.11.003
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Here we report on the construction and evaluation of a bifunctional combined aptamer (BCA) that consists of a DNA streptavidin-binding aptamer (SBA), a DNA thrombin-binding aptamer (TBA) and a fluorophore. The BCA adopts a new conformation that is very different from simply linking the conformations of the two individual aptamers together, so that it does not bind to streptavidin in the absence of thrombin. Binding of this novel DNA aptamer to streptavidin is triggered by the thrombin binding and depends on the concentration of thrombin. Meanwhile, fluorescence from the streptavidin captured BCA reflects the quantity of the target molecule in the sample. This aptamer combination strategy based on the SBA holds good potential for applications in simultaneous detection and separation of targets of aptamers or certain DNA and RNA targets. (c) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:1487 / 1492
页数:6
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