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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

    Year: 2019

    Journal: Microchim. Acta, Volume 186, AUG

    Authors: Diaz-Betancor, Zeneida; Banuls, Maria-Jose; Javier Sanza, Francisco; Casquel, Rafael; Fe Laguna, Maria; Holgado, Miguel; Puchades, Rosa; Maquieira, Angel

    Organizations: FEDEREuropean Commission [CTQ/2016/75749-R, TEC/2017/84846-R]; MINECO [CTQ/2016/75749-R, TEC/2017/84846-R]

    Keywords: Thiol-acrylate photocoupling; SU-8; Blu-ray disc; Interferometric detection; C-reactive protein; Nucleic acid; Bovine serum albumin; UV-light

    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 (254nm). 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 (lambda(em) 635nm) 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 30ngmL(-1) and 2pgmL(-1), respectively.
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