title: Small molecule aptamer assays based on fluorescence anisotropy signal-enhancer oligonucleotides.
abstract: Herein, we design novel fluorescence anisotropy (FA) aptamer sensing platforms dedicated to small molecule detection. The assay strategy relied on enhanced fluctuations of segmental motion dynamics of the aptamer tracer mediated by an unlabelled, partially complementary oligonucleotide. The signal-enhancer oligonucleotide (SEO) essentially served as a free probe fraction revealer. By targeting specific regions of the signalling functional nucleic acid, the SEO binding to the unbound aptamer triggered perturbations of both the internal DNA flexibility and the localized dye environment upon the free probe to duplex structure transition. This potentiating effect determined increased FA variations between the duplex and target bound states of the aptameric probe. FA assay responses were obtained with both pre-structured (adenosine) and unstructured (tyrosinamide) aptamers and with dyes of different photochemical properties (fluorescein and texas red). The multiplexed analysis ability was further demonstrated through the simultaneous multicolour detection of the two small targets. The FA method appears to be especially simple, sensitive and widely applicable.
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Perrier
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Prisca
authors.initials: P
authors.lastname: Bouilloud
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Gisella
authors.initials: G
authors.lastname: De Oliveira Coelho
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Mickael
authors.initials: M
authors.lastname: Henry
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France. Electronic address: eric.peyrin@ujf-grenoble.fr.
authors.forename: Eric
authors.initials: E
authors.lastname: Peyrin
mesh_terms: D000241:Adenosine / Q000032:analysis*; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D019793:Fluorescein / Q000737:chemistry; D005454:Fluorescence Polarization / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; C036387:tyrosinamide; D014443:Tyrosine; D000241:Adenosine; D019793:Fluorescein
keywords: Aptamer; Fluorescence anisotropy; Multiplexed analysis; Segmental motion; Signal enhancement; Small target