Lian Zewei, Chi Jimei, Yang Xu, Cheng Lijun, Xie Daixi, Tan Zhiyu, Chen Sisi, Yun Yang, Yibulayimu Yisilamu, Wu Wei, Song Yanlin, Su Meng
Key Laboratory of Green Printing, Institute of Chemistry, Chinese Academy of Sciences/Beijing Engineering Research Center of Nanomaterials for Green Printing Technology, Beijing National Laboratory for Molecular Sciences, Beijing, China.
University of Chinese Academy of Sciences, Beijing, China.
Nat Protoc. 2025 Jul 24. doi: 10.1038/s41596-025-01215-y.
There is an urgent requirement to improve accessibility to diagnostic tools in remote areas. This requires assays that are easy to use, are cost-effective and produce rapid results. Important public health applications include early disease diagnosis, real-time monitoring, epidemic control and medical cost control. This protocol describes the fabrication of all-printed photonic crystal (PC) biochips for point-of-care testing of biomarkers. The photonic crystal material is prepared by the self-assembly of latex nanospheres that are printed onto a polyethylene terephthalate substrate. Photonic crystals composed of latex nanospheres of different sizes enhance the fluorescent signal emitted at different wavelengths, resulting in remarkably higher detection sensitivity. PC microarrays enable mass-printed preparation (up to 2,700 pieces can be printed per hour by one printer), and each microarray can be stored for a long time (>6 months) after heating. Biomarker specificity is achieved by the biofunctionalization of the nanospheres, for example, attaching capture antibodies. The detection involves the use of a fluorescently labeled detection antibody and a simple point-of-care detection device. This universal approach can be applied to the detection of many biomarkers, and the simultaneous detection of multiple biomarkers is also possible. Here we demonstrate describing how to prepare a chip that can be used to detect three inflammatory biomarkers in 10 min at low sample volumes at a cost of less than 3 CNY (~US$0.41) per PC codetection biochip. The biofunctionalization process including capture antibody coupling and blocking takes 3-4 h, and the detection process takes 20 min.
迫切需要提高偏远地区诊断工具的可及性。这就需要易于使用、具有成本效益且能快速出结果的检测方法。重要的公共卫生应用包括疾病早期诊断、实时监测、疫情控制和医疗成本控制。本方案描述了用于生物标志物即时检测的全印刷光子晶体（PC）生物芯片的制备。光子晶体材料通过乳胶纳米球的自组装制备，然后印刷到聚对苯二甲酸乙二酯基板上。由不同大小乳胶纳米球组成的光子晶体增强了在不同波长发射的荧光信号，从而显著提高检测灵敏度。PC微阵列能够进行大规模印刷制备（一台打印机每小时可印刷多达2700片），并且每个微阵列在加热后可长期保存（>6个月）。通过纳米球的生物功能化实现生物标志物特异性，例如连接捕获抗体。检测使用荧光标记的检测抗体和简单的即时检测装置。这种通用方法可应用于多种生物标志物的检测，同时检测多种生物标志物也是可行的。在此，我们展示了如何制备一种芯片，该芯片可在低样本量下于10分钟内检测三种炎症生物标志物，每个PC共检测生物芯片成本低于3元人民币（约0.41美元）。包括捕获抗体偶联和封闭在内的生物功能化过程需要3 - 4小时，检测过程需要20分钟。
