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Phosphorylcholine-based hydrogel for immobilization of biomolecules. Application to fluorometric microarrays for use in hybridization assays and immunoassays, and nanophotonic biosensing.
Díaz-Betancor, Zeneida; Bañuls, María-José; Sanza, Francisco Javier; Casquel, Rafael; Laguna, Maria Fe; Holgado, Miguel; Puchades, Rosa; Maquieira, Ángel.
Afiliação
  • Díaz-Betancor Z; IDM, Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico, Universitat Politècnica de València, Universitat de València, Camino de Vera s/n, 46022, Valencia, Spain.
  • Bañuls MJ; IDM, Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico, Universitat Politècnica de València, Universitat de València, Camino de Vera s/n, 46022, Valencia, Spain. mbpolo@upvnet.upv.es.
  • Sanza FJ; Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022, Valencia, Spain. mbpolo@upvnet.upv.es.
  • Casquel R; Center for Biomedical Technology, Optics, Photonics and Biophotonics Lab, Universidad Politécnica de Madrid, Campus Montegancedo, 28223, Madrid, Spain.
  • Laguna MF; Bio Optical Detection S.L., Isaac Peral 1, 28840, Madrid, Spain.
  • Holgado M; Center for Biomedical Technology, Optics, Photonics and Biophotonics Lab, Universidad Politécnica de Madrid, Campus Montegancedo, 28223, Madrid, Spain.
  • Puchades R; Center for Biomedical Technology, Optics, Photonics and Biophotonics Lab, Universidad Politécnica de Madrid, Campus Montegancedo, 28223, Madrid, Spain.
  • Maquieira Á; Center for Biomedical Technology, Optics, Photonics and Biophotonics Lab, Universidad Politécnica de Madrid, Campus Montegancedo, 28223, Madrid, Spain.
Mikrochim Acta ; 186(8): 570, 2019 07 24.
Article em En | MEDLINE | ID: mdl-31338609
An approach is presented for covalent immobilization of biomolecules on an acrylate phosphorylcholine hydrogel. The immobilization and the hydrogel formation take place simultaneously by a thiol-acrylate coupling reaction, induced by UV-light (254 nm). The hydrogel is prepared on two polymeric surfaces (the HardCoat protective layer of Blu-Ray discs, and SU-8) and applied to fluorescence microarray and label-free interferometric detection. For the first, Cy5 labeled analytes are used (λem 635 nm) and, for the second, a periodic array of high-aspect ratio nanopillars detects unlabeled analytes by interferometry. Bioavailability of the immobilized probes is demonstrated in labeled assays; for the case of oligonucleotides by discriminating single nucleotide polymorphisms, and, for the case of antibodies, by BSA immunorecognition. The raw hydrogel is employed to detect human C-reactive protein, in both labeled and non-labeled assay formats, with sensitivities of 30 ng·mL-1 and 2 pg·mL-1, respectively. Graphical abstract Schematic presentation of the phosphorylcholine (MPC) hydrogel preparation onto BluRay disc and SU-8 nanopillars to perform fluorescence and label-free interferometric detection, respectively. It selectively detects C-reactive protein (CRP), but it can covalently immobilize antibodies or nucleid acid probes to detect other analytes.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Imunoensaio / Técnicas Biossensoriais / Hidrogéis / Análise em Microsséries / Fluorometria / Hibridização de Ácido Nucleico Limite: Humans Idioma: En Ano de publicação: 2019 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Imunoensaio / Técnicas Biossensoriais / Hidrogéis / Análise em Microsséries / Fluorometria / Hibridização de Ácido Nucleico Limite: Humans Idioma: En Ano de publicação: 2019 Tipo de documento: Article