基于纳米孔技术的肺癌相关活性分子检测及电化学分析机制探索

The expression of lung cancer-related bioactive molecules is closely related to the progression of lung cancer. Development of a method for sensitive and selective detection of bioactive molecules is of great significance. Nanopore-based single-molecule technology has the advantages of being rapid, low-cost, and label-free, providing a new platform for the detection of biomolecules. This project aims to detect the nucleic acid and protein markers associated with lung cancer. Bioprobes with high specificity to the target molecules will be designed. A series of glass nanopores can be fabricated by laser pulling technique, and functional modification will be performed. Aerolysin is to be used to form the biological nanopore. Based on that, we would build nanopore sensors for the detection of various lung cancer biomarkers. Besides, signal amplification strategy will be employed to improve the sensitivity. The specificity is expected to be improved by the simultaneous determination of various active molecules. Furthermore, nanopore electrochemical analysis technology will be combined with dark field imaging technique. In this way, we would monitor the electron transfer process in the nanopore interface, and explore the mechanism of nanopore electrochemical analysis.

肺癌相关活性分子的表达量与肺癌的发生发展密切相关，发展一种灵敏度高、特异性强的活性分子检测技术具有重要意义。纳米孔单分子分析技术具有快速、低成本、无需荧光标记等优点，为生物分子的检测提供了一个新平台。本项目拟以肺癌相关的核酸和蛋白质标志物为检测对象，设计特异性识别目标物的功能化生物探针；利用激光拉制技术制备一系列玻璃纳米孔道，进行功能化修饰，同时利用气单胞菌溶素（Aerolysin）自组装形成生物纳米孔道，建立肺癌相关活性分子的纳米孔传感分析体系；引入信号放大策略提高灵敏度，通过多种活性分子的同时测定提高特异性；进一步将纳米孔电化学分析技术与暗场显微成像相结合，监测纳米孔道界面的电子传递过程，探索纳米孔电化学分析机制。

肺癌相关活性分子的表达量与肺癌的发生发展密切相关，发展一种灵敏度高、特异性强的活性分子检测技术具有重要意义。纳米孔单分子分析技术具有快速、低成本、无需荧光标记等优点，为生物分子的检测提供了一个新平台。本项目以核酸和蛋白质等生物标志物为检测对象，分别以DNA和多肽为结构单元，结合利用纳米金、磁性纳米粒子等材料的优异性能，开发特异性识别目标物的功能化生物探针，包括分叉DNA探针、双链DNA纳米探针、多肽探针等。利用气单胞菌溶素（aerolysin）和α-溶血素分别自组装形成生物纳米孔道，发展了一系列纳米孔传感新方法，实现了多种肺癌相关活性分子的检测；纳米孔结合分子动力学模拟，研究序列改变对DNA二级结构i-motif稳定性的影响，实现核酸结构在单分子水平的实时监测；进一步，通过点击反应引入DNA分支设计条码探针，结合纳米孔传感界面，实现了多种蛋白质的联合检测和精确区分，该方法具有通用性，改变碱基种类、序列长度可同时分析更多目标物。将所建立的方法应用于血清样品分析，表现出良好的灵敏度和选择性，在生化分析中具有较好的应用前景。