基于中空通道纸芯片的自推进分离捕获循环肿瘤细胞及原位分析

The detection of circulating tumour cells from peripheral blood is the most innovative and potential tumor diagnosis method, which can be regard as a non-invasion ‘liquid biopsy’ of the tumor. However, isolation, detection and characterization of circulating tumour cells has been technically challenging owing to their extremely low number in the bloodstream. In this project, hollow channel is introduced into microfluidic paper chip to develop the multiple seperation and enrichment paper chip based on filter membrane at the entrance, characteristic of cell biology in hollow channel and spatial effect of hollow channel. To further improve cell-capture performance, nanomaterials modified hollow channel affect the type of flow, which can increase the probability of cells and recognizer molecule. The hollow channel and the detection zone is loaded cells recognizer molecule, which realize the negative isolation and positive enrichment, respectively. Coralline paper electrode is prepared based on the porous property and chemical growth method, which is used to determine cell. Some nanocomposites probes are prepared, the signal is magnified via biotechnology and nanocomposites probes. The platform of hollow channel microfluidic paper chip is integrated, which is used to study tumor cells, early diagnosis of cancer, drug screening and the application of whole blood.

外周血中循环肿瘤细胞检测列为最具创新性和转化潜力的肿瘤诊断新方法，它是一种非侵袭性“液体活检”方式，但其存在数量低、分离富集检测困难等问题。本项目引入中空通道至微流控纸芯片，开发通道底部半亲水和半疏水功能实现自推进分离捕获循环肿瘤细胞；采用进样区域置入滤膜过滤分离、中空通道利用细胞生物学特性分离、通道空间效应分离相结合制备细胞的多重分离富集纸芯片。中空通道修饰纳米材料实现液体流动方式可控，增加细胞与识别分子接触几率。筛选组合细胞识别分子负载中空通道与检测区域，中空通道与检测区域分别进行正常细胞阴性分选和肿瘤细胞阳性富集。基于纸纤维多孔性、化学生长珊瑚状金属纸电极，提高电极导电性及易光生载流子传输，应用纳米复合探针及分子生物学技术信号放大，实现细胞测定。集成中空通道纸芯片平台，利用平台对肿瘤细胞进行研究、癌症早期诊断、药物筛选及在血液样本中的应用。

外周血循环肿瘤细胞（CTCs）的检测对肿瘤的早期诊断、判断患者预后及疗效评估等都具有重要意义。由于CTCs在外周血中的数量极少, 因而对其进行分离、富集，信号放大是高灵敏的CTCs检测新方法需要解决的重要问题。按照项目计划书的目标和计划，执行过程中完成了项目计划书中所列目标。首先，利用纸折叠或者多层纸叠加制备微流控纸芯片，实现了滤膜过滤分离、细胞生物学特性中空通道分离及利用通道空间效应对CTCs的多重分离和富集；其次，电极纸纤维表面制备了导电性好及易光生载流子传输的功能材料，实现了电化学和光致电化学检测的高性能传感界面；再次，利用纳米材料和生物技术相结合开发出了多种具有靶向识别和信号放大的纳米复合探针；然后通过分子生物学过程，建立多途径信号放大策略，提高检测的准确度和灵敏度；最后，开发出集分离富集检测CTCs于一体的微流控纸芯片平台，实现了循环肿瘤细胞的快速检测。围绕该项目获国家发明专利12项、发表SCI收录论文30余篇、山东省自然科学一等奖1项、中国分析测试协会科学技术奖二等奖1项、培养博士2名、硕士8名、获得山东省高等学校青年创新团队。