title: Fluorescence anisotropy-based structure-switching aptamer assay using a peptide nucleic acid (PNA) probe.
abstract: This study describes for the first time the feasibility of using peptide nucleic acids (PNAs) as an alternative to the DNA probes in structure-switching aptamer fluorescence polarisation assays. The effects of experimental parameters such as the length of the PNA strand, the nature of dye and the buffer conditions on the assay performances are first explored using two different methodologies based on the competition between the PNA/aptamer hydribridisation and the target/aptamer complexation. D-ATP can be detected from 1 to 25 μM in a linear range and a detection limit (LOD) of 3 μM can be reached. For this target, this lowers by a factor >5 the LOD reported with conventional DNA-based fluorescent structure switching aptamer-based assays and by a factor 3 the LOD observed with non-competitive fluorescent sensing platform indicating the usefulness of the PNA-based approach.
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Emma
authors.initials: E
authors.lastname: Goux
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Quentin
authors.initials: Q
authors.lastname: Lespinasse
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France. Electronic address: valerie.guieu@ujf-grenoble.fr.
authors.forename: Valérie
authors.initials: V
authors.lastname: Guieu
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: Corinne
authors.initials: C
authors.lastname: Ravelet
authors.affiliation: Département de Pharmacochimie Moléculaire, Université Grenoble Alpes, UMR 5063 CNRS, ICMG FR 2607, Campus universitaire, Saint-Martin d'Hères, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Fiore
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: D000255:Adenosine Triphosphate / Q000032:analysis*; D052157:Aptamers, Nucleotide / Q000737:chemistry; D015374:Biosensing Techniques; D005454:Fluorescence Polarization; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D020135:Peptide Nucleic Acids / Q000737:chemistry*
chemical_list: D052157:Aptamers, Nucleotide; D020135:Peptide Nucleic Acids; D000255:Adenosine Triphosphate
keywords: Aptamer-based assay; Fluorescence anisotropy; Peptide nucleic acid; Structure-switching