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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

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Abstract

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.

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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Acknowledgements

This research was funded by FEDER and the MINECO projects CTQ/2016/75749-R and TEC/2017/84846-R. The authors acknowledge the contribution of the Bc’s D students Alejandra Vargas and Ismael Guerrero for the time spent at our lab and for their enthusiastic ideas on the possibilities of MPC hydrogel.

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Authors and Affiliations

  1. 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

    Zeneida Díaz-Betancor, María-José Bañuls, Rosa Puchades & Ángel Maquieira

  2. Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022, Valencia, Spain

    María-José Bañuls, Rosa Puchades & Ángel Maquieira

  3. Center for Biomedical Technology, Optics, Photonics and Biophotonics Lab, Universidad Politécnica de Madrid, Campus Montegancedo, 28223, Madrid, Spain

    Francisco Javier Sanza, Rafael Casquel, Maria Fe Laguna & Miguel Holgado

  4. Bio Optical Detection S.L., Isaac Peral 1, 28840, Madrid, Spain

    Francisco Javier Sanza

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  1. Zeneida Díaz-Betancor
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Díaz-Betancor, Z., Bañuls, MJ., Sanza, F.J. et al. Phosphorylcholine-based hydrogel for immobilization of biomolecules. Application to fluorometric microarrays for use in hybridization assays and immunoassays, and nanophotonic biosensing. Microchim Acta 186, 570 (2019). https://doi.org/10.1007/s00604-019-3691-3

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