Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Universitat Politécnica de Valéncia, Universitat de Valéncia, Camino de Vera s/n, 46022 Valencia, Spain.
Departamento de Química, Universitat Politécnica de Valéncia, Camino de Vera s/n, 46022 Valencia, Spain.
Biosensors (Basel). 2023 Feb 23;13(3):312. doi: 10.3390/bios13030312.
The present research is focused on the development of a biofunctionalized hydrogel with a surface diffractive micropattern as a label-free biosensing platform. The biosensors described in this paper were fabricated with a holographic recording of polyethylene terephthalate (PET) surface micro-structures, which were then transferred into a hydrogel material. Acrylamide-based hydrogels were obtained with free radical polymerization, and propargyl acrylate was added as a comonomer, which allowed for covalent immobilization of thiolated oligonucleotide probes into the hydrogel network, via thiol-yne photoclick chemistry. The comonomer was shown to significantly contribute to the immobilization of the probes based on fluorescence imaging. Two different immobilization approaches were demonstrated: during or after hydrogel synthesis. The second approach showed better loading capacity of the bioreceptor groups. Diffraction efficiency measurements of hydrogel gratings at 532 nm showed a selective response reaching a limit of detection in the complementary DNA strand of 2.47 µM. The label-free biosensor as designed could significantly contribute to direct and accurate analysis in medical diagnosis as it is cheap, easy to fabricate, and works without the need for further reagents.
本研究专注于开发具有表面衍射微图案的生物功能化水凝胶，作为无标记生物传感平台。本文中描述的生物传感器是通过对聚对苯二甲酸乙二醇酯 (PET) 表面微结构进行全息记录来制造的，然后将其转移到水凝胶材料中。通过自由基聚合获得丙烯酰胺基水凝胶，并添加炔丙基丙烯酸酯作为共聚单体，通过硫醇-炔点击化学将巯基化寡核苷酸探针共价固定在水凝胶网络中。共单体的存在基于荧光成像显示出对探针的固定有显著贡献。展示了两种不同的固定方法：在水凝胶合成过程中或之后。第二种方法显示出更好的生物受体基团的负载能力。在 532nm 处对水凝胶光栅的衍射效率测量显示出选择性响应，互补 DNA 链的检测限达到 2.47µM。设计的无标记生物传感器可以为医学诊断中的直接和准确分析做出重要贡献，因为它价格便宜、易于制造，并且无需进一步的试剂即可工作。
