Noncompetitive fluorescence polarization aptamer-based assay for small molecule detection.
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.
Ruta
Josephine
J
Département de Pharmacochimie Moléculaire UMR 5063, Institut de Chimie Moléculaire de Grenoble FR 2607, CNRS-Université Grenoble I (Joseph Fourier), 38041 Grenoble cedex 9, France.
Perrier
Sandrine
S
Ravelet
Corinne
C
Fize
Jennifer
J
Peyrin
Eric
E
D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D015374:Biosensing Techniques / Q000379:methods*; D005454:Fluorescence Polarization / Q000379:methods*; D005456:Fluorescent Dyes; D009690:Nucleic Acid Conformation; D012680:Sensitivity and Specificity; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism
D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; C036387:tyrosinamide; D014443:Tyrosine