Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Universitat Politècnica de València, Universitat de València, Valencia, Spain.
Departamento de Química, Universitat Politècnica de València, Valencia, Spain.
Methods Mol Biol. 2022;2393:57-72. doi: 10.1007/978-1-0716-1803-5_4.
We have developed large-scale one-dimensional photonic crystals from standard recordable Blu-ray disks, tailored to sense unlabeled biorecognition events on their surface. These materials rely on coating, with layers of 80 nm of titanium oxide, nanogrooved polycarbonate plates obtained from regular disks. As a result, they present guided-mode resonances that we have demonstrated that can be exploited to quantify biorecognition events by means of the bandgap positions in the transmission spectra. These photonic crystals have displayed well-correlated dose-response curves in immunoassays to quantify IgGs, C-reactive protein, and lactate dehydrogenase. The detection limit reached is 16 ng/mL, 2μg/mL, and 18 ng/mL, respectively. Herein we describe the experimental procedures and methods to fabricate and functionalize these photonic crystals, perform immunoassays on them, set up an optical system to measure their response, and process the resulting data to perform bioanalytical determinations in label-free format.
我们已经从标准可记录蓝光光盘中开发出了大规模的一维光子晶体，可用于感测其表面上未标记的生物识别事件。这些材料依赖于涂覆有 80nm 厚的氧化钛层，以及从普通光盘获得的纳米槽聚碳酸酯板。结果，它们呈现出导模共振，我们已经证明可以利用传输光谱中的带隙位置来定量生物识别事件。这些光子晶体在免疫测定中显示出良好的剂量响应曲线，可以定量测定 IgG、C 反应蛋白和乳酸脱氢酶。分别达到的检测限为 16ng/mL、2μg/mL 和 18ng/mL。本文描述了制造和功能化这些光子晶体、在其上进行免疫测定、设置光学系统来测量其响应以及处理所得数据以进行无标记格式的生物分析测定的实验程序和方法。
