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.
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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.
The author(s) declare that they have no competing interests.
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Díaz-Betancor, Z., Bañuls, M., 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
- Thiol-acrylate photocoupling
- Blu-ray disc
- Interferometric detection
- C-reactive protein
- Nucleic acid
- Bovine serum albumin