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.
Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022, Valencia, Spain.
Mikrochim Acta. 2019 Jul 24;186(8):570. doi: 10.1007/s00604-019-3691-3.
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 (λ 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 and 2 pg·mL, 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.
介绍了一种将生物分子共价固定在丙烯酰基磷酸胆碱水凝胶上的方法。通过硫醇-丙烯酯偶联反应,在紫外光(254nm)的诱导下同时进行固定和水凝胶的形成。该水凝胶在两个聚合物表面(蓝光光盘的 HardCoat 保护层和 SU-8)上制备,并应用于荧光微阵列和无标记干涉检测。对于前者,使用 Cy5 标记的分析物(λ 635nm),对于后者,通过干涉测量检测未标记的分析物的高纵横比纳米柱周期性阵列。在标记测定中证明了固定探针的生物可用性;对于寡核苷酸,通过区分单核苷酸多态性,对于抗体,通过 BSA 免疫识别。原始水凝胶用于在标记和非标记测定格式中检测人 C 反应蛋白,分别具有 30ng·mL 和 2pg·mL 的灵敏度。