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Noncompetitive Fluorescence Polarization Aptamer-based Assay for Small Molecule Detection

2009; American Chemical Society; Volume: 81; Issue: 17 Linguagem: Inglês

10.1021/ac9014512

1520-6882

Joséphine Ruta, Sandrine Perrier, Corinne Ravelet, Jennifer Fize, Éric Peyrin,

DNA and Nucleic Acid Chemistry

In this paper, a new fluorescence polarization (FP) assay strategy is described reporting the first demonstration of a noncompetitive FP technique dedicated to the small molecule sensing. This approach was based on the unique induced-fit binding mechanism of nucleic acid aptamers which was exploited to convert the small target binding event into a detectable fluorescence anisotropy signal. An anti-L-tyrosinamide DNA aptamer, labeled by a single fluorescent dye at its extremity, was employed as a model functional nucleic acid probe. The DNA conformational change generated by the L-tyrosinamide binding was able to induce a significant increase in the fluorescence anisotropy signal. The method allowed enantioselective sensing of tyrosinamide and analysis in practical samples. The methodology was also applied to the L-argininamide detection, suggesting the potential generalizability of the direct FP-based strategy. Such aptamer-based assay appeared to be a sensitive analytical system of remarkable simplicity and ease of use.