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Non-SELEX isolation of DNA aptamers for the homogeneous-phase fluorescence anisotropy sensing of tau Proteins.
Lisi, Samuele; Fiore, Emmanuelle; Scarano, Simona; Pascale, Emanuela; Boehman, Yannik; Ducongé, Frederic; Chierici, Sabine; Minunni, Maria; Peyrin, Eric; Ravelet, Corinne.
Afiliación
  • Lisi S; Univ. Grenoble Alpes, DPM UMR 5063, CNRS, F-38041 Grenoble, France; Department of Chemistry 'Ugo Schiff', Via della Lastruccia 3-13, Sesto Fiorentino, 50019, Firenze, Italy.
  • Fiore E; Univ. Grenoble Alpes, DPM UMR 5063, CNRS, F-38041 Grenoble, France.
  • Scarano S; Department of Chemistry 'Ugo Schiff', Via della Lastruccia 3-13, Sesto Fiorentino, 50019, Firenze, Italy.
  • Pascale E; Department of Chemistry 'Ugo Schiff', Via della Lastruccia 3-13, Sesto Fiorentino, 50019, Firenze, Italy.
  • Boehman Y; Unité des virus émergents, UMR IRD 190 EPV, Faculté de Médecine-Timone, 27 boulevard Jean-Moulin, 13385 Marseille Cedex 5, France.
  • Ducongé F; CEA, Fundamental Research Division (DRF), Institute of Biology François Jacob (Jacob), Molecular Imaging Research Center, Neurodegenerative Diseases Laboratory, CNRS UMR 9199, Paris-Saclay University, Fontenay aux Roses, France.
  • Chierici S; Univ. Grenoble Alpes, DCM UMR 5250, CNRS, F-38041 Grenoble, France.
  • Minunni M; Department of Chemistry 'Ugo Schiff', Via della Lastruccia 3-13, Sesto Fiorentino, 50019, Firenze, Italy.
  • Peyrin E; Univ. Grenoble Alpes, DPM UMR 5063, CNRS, F-38041 Grenoble, France. Electronic address: eric.peyrin@univ-grenoble-alpes.fr.
  • Ravelet C; Univ. Grenoble Alpes, DPM UMR 5063, CNRS, F-38041 Grenoble, France. Electronic address: corinne.ravelet@ujf-grenoble.fr.
Anal Chim Acta ; 1038: 173-181, 2018 Dec 14.
Article en En | MEDLINE | ID: mdl-30278900
Herein, we report for the first time the isolation of DNA aptamers directed against the whole tau protein, an important Alzheimer's disease (AD) biomarker. Non-SELEX approach based on the capillary electrophoresis partitioning technique was employed to isolate a high-affinity DNA sequence pool towards the target in only three rounds and one working day. High-throughput sequencing was next performed and the recognition ability of five selected aptamers was preliminary evaluated by surface plasmon resonance using the protein target immobilized on the chip. Finally, the analytical potential of the most affine aptamer was demonstrated through the design of a homogeneous-phase fluorescence anisotropy assay. This DNA aptamer was found to be able to recognize not only the whole τ-441 but also the τ-381, τ-352, τ-383 isoforms. The sensing platform allowed the determination of these four targets with a detection limit of 28 nM, 3.2 nM, 6.3 nM and 22 nM, respectively.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Proteínas tau / Aptámeros de Nucleótidos / Polarización de Fluorescencia Límite: Humans Idioma: En Revista: Anal Chim Acta Año: 2018 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Proteínas tau / Aptámeros de Nucleótidos / Polarización de Fluorescencia Límite: Humans Idioma: En Revista: Anal Chim Acta Año: 2018 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Países Bajos