新型自供能细胞传感平台构建及循环肿瘤细胞检测研究

Circulating tumor cells (CTCs) in peripheral blood play crucial roles in cancer metastasis process, therefore, there is an urgent need to develop ultrasensitive testing approachs, even at the single-cell level, for early cancer diagnosis and treatment. Self-powered biosensors have the merits of needing no external power sources, simple fabrication process and strong anti-interference ability, making them promising candidates for CTCs detection in human disease diagnosis. The core technique of self-powered biosensors is the fabrication of the high-performance enzymic biofuel cell (EBFC). Meanwhile, DNA amplification techniques have been widly used for improving the sensitivity of the biosensors. Thus, this project is proposed to develop a novel self-powered cytosensing strategy for the detection of CTCs in peripheral blood based on dual-signal amplification of high-performance EBFC and DNA amplification techniques. A new self-powered cytosensing platform will be built in order to realize simple, fast, highly selective and ultrasensitive detection of CTCs. Furthermore, this cytosensor does not need external voltage, which efficiently eliminates the non-specific redox of the electro-active interferences, improves the anti-fouling ability of the cytosensor, and facilitates the detection of CTCs in peripheral blood samples. This project would provide some new methods and techniques for the early cancer diagnosis. Some innovative research results are also expected with the execution of the current project.

循环肿瘤细胞经血道转移可导致肿瘤扩散，实现肿瘤外周血中低丰度循环肿瘤细胞的超灵敏检测，甚至单细胞水平检测，在癌症诊断和治疗中至关重要。基于酶生物燃料电池的自供能生物传感器无需额外供电设备、测试体系简单、抗干扰能力强、可实现目标物简单、快速、实时监测。高性能酶生物燃料电池是构建高灵敏自供能生物传感器的核心，同时DNA放大技术能有效提高生物传感器灵敏度。本项目拟基于高性能酶生物燃料电池与新型DNA放大技术，创立双重信号放大的自供能细胞传感新模式，构建简单、快速、高选择性、超灵敏检测肿瘤细胞的传感平台。自供能检测体系无需额外施加电压，能有效避免外周血中氧化还原组分干扰，实现外周血中低丰度循环肿瘤细胞的“精准”检测。本项目的顺利实施，以期为癌症早期诊断提供新技术和新方法，有望获得具有一定特色的创新性研究成果。

发展高灵敏、特异性检测循环肿瘤细胞、疾病标志物的生物传感技术，实现快速检测对于重大疾病的临床早期诊断、治疗具有十分重要的研究意义。本项目开展了循环肿瘤细胞超灵敏检测新方法研究，建立了光诱导的无阳极酶、无隔膜、无媒介体的自供能循环肿瘤细胞传感平台，构建了免标记、无酶、超灵敏均相电化学比率传感器检测循环肿瘤细胞。开展了肿瘤标志物传感新方法研究，基于硫代磷酸化作用介导的自供能生物传感器超灵敏地检测蛋白激酶A活性及抑制剂。集成酶生物燃料电池（EBFC）与DNA放大策略，构建了自供能生物传感器用于单核苷酸多态性的超灵敏、高特异性检测。集EBFCs与均相DNA电化学于一体，构建了一种新型的均相自供能生物传感策略，实现了microRNA的超灵敏检测。开展了自供能生物传感应用于食品安全方面的检测研究，EBFC与DNA生物共轭体协同作用，实现了牛奶中的抗生素（氨苄西林）药物残留的超灵敏检测。基于目标物诱导阳极燃料分子释放的设计策略，构建了基于EBFC的自供能均相免疫传感器，并用于超灵敏检测牛奶中的三聚氰胺。光电酶生物燃料电池构筑了一种光驱动的自供能生物传感方法实现了有机磷农药的超灵敏的检测。开展了无酶生物燃料电池（NEFCs）传感研究，利用电嫁接有机铜络合物阴极和多孔金纳米粒子阳极构筑了高性能无酶生物燃料电池，并开展了自供能生物传感研究，实现了对焦磷酸盐的快速，高效，灵敏检测。.在国家自然科学基金的资助下，已发表 SCI 收录论文 13 篇，其中影响因子均大于5.0。培养了6名硕士专业人才。项目主持人获山东省省属优青。

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