Fluorescence polarization biosensor based on an aptamer enzymatic cleavage protection strategy
| Autor: | Anthony Kidd, Corinne Ravelet, Eric Peyrin, Valérie Guieu, Sandrine Perrier |
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| Rok vydání: | 2011 |
| Předmět: |
chemistry.chemical_classification
Adenosine Aptamer Fluorescence Polarization Model system Biosensing Techniques Aptamers Nucleotide Ochratoxins Biochemistry Analytical Chemistry Phosphodiesterase I chemistry.chemical_compound Enzyme chemistry Biophysics Biosensor DNA Fluorescence anisotropy Fluorescent Dyes |
| Zdroj: | Analytical and Bioanalytical Chemistry. 401:3229-3234 |
| ISSN: | 1618-2650 1618-2642 |
| DOI: | 10.1007/s00216-011-5434-0 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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