Noncompetitive Fluorescence Polarization Aptamer-based Assay for Small Molecule Detection

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• 作者：Josephine Ruta ; Sandrine Perrier ; Corinne Ravelet ; Jennifer Fize ; Eric Peyrin
• 刊名：Analytical Chemistry
• 出版年：2009
• 出版时间：September 1, 2009
• 年：2009
• 卷：81
• 期：17
• 页码：7468-7473
• 全文大小：183K
• 年卷期：v.81,no.17(September 1, 2009)
• ISSN：1520-6882

文摘

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.