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Aptamer-based label-free method for hemin recognition and DNA assay by capillary electrophoresis with chemiluminescence detection
被引:57
作者:
Li, Tao
Li, Bingling
Dong, Shaojun
[1
]
机构:
[1] Chinese Acad Sci, Changchun Inst Appl Chem, State Key Lab Electroanalyt Chem, Changchun 130022, Jilin, Peoples R China
[2] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
基金:
中国国家自然科学基金;
关键词:
aptamer;
hemin;
DNA;
molecular recognition;
capillary electrophoresis;
D O I:
10.1007/s00216-007-1487-5
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
An aptamer-based label-free approach to hemin recognition and DNA assay using capillary electrophoresis with chemiluminescence detection is introduced here. Two guanine-rich DNA aptamers were used as the recognition element and target DNA, respectively. In the presence of potassium ions, the two aptamers folded into the G-quartet structures, binding hemin with high specificity and affinity. Based on the G-quartet-hemin interactions, the ligand molecule was specifically recognized with a K (d)approximate to 73 nM, and the target DNA could be detected at 0.1 mu M. In phosphate buffer of pH 11.0, hemin catalyzed the H2O2-mediated oxidation of luminol to generate strong chemiluminescence signal; thus the target molecule itself served as an indicator for the molecule-aptamer interaction, which made the labeling and/or modification of aptamers or target molecules unnecessary. This label-free method for molecular recognition and DNA detection is therefore simple, easy, and effective.
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页码:887 / 893
页数:7
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