期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 401, 期 10, 页码 3229-3234出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-011-5434-0
关键词
Fluorescence polarization; Aptamer; Enzymatic cleavage protection; Small analyte; Sensing
资金
- SEST-Micraptox [2007-013-01]
A novel fluorescence polarization (FP) aptasensing platform based on target-induced aptamer enzymatic cleavage protection is reported. The method relies on the FP analysis of the phosphodiesterase I mediated size variation of a dye-labeled aptamer. The tyrosinamide/antityrosinamide DNA aptamer couple was firstly tested as a model system to establish the proof-of-concept. In the absence of the target, the labeled aptamer was enzymatically cleaved into small DNA fragments, leading to a low FP signal. Upon tyrosinamide binding, the DNA substrate was partially protected against the enzymatic attack, leading to an increase in the fluorescence anisotropy response as a result of the higher average molecular volume of the weakly digested probe. The method was subsequently applied to two other systems, i.e., for the detection of ochratoxin A and adenosine. Such an approach was found to combine simplicity and general applicability features.
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