靶向肽纸芯片甄别检测高侵袭恶性循环肿瘤细胞的新方法

The demand for the analysis of the abundance and activity of mesenchyme-like marker protein between highly invasive vicious circulating tumor cells (CTCs) is continuously increasing. In this project, hollow channel is fabricated on the microfluidic lab-on-paper to improve the efficiency of the separation and concentration of CTCs. Paper-based electronic devices are utilized for the fabrication of electronic catalytic reactor, self-power device and signal storage device, to realize the collection, conversion and modulation between optical and electronic signals. The marker protein for epithelial–mesenchymal transitions of highly invasive vicious CTCs in structural biology is to be determinedand the peptide with specific binding site and conformation will be screened by library screening technology. Superior peptide motif with the prominent function of molecular recognition and signal amplification is to be developed via algorithm optimization, artificial modification and functional motif combination. Hydrophilic or amphiphilic materials are to be modified onto the surface of electrodes to fabricate anti-pollution sensor interfaces as well as uniformly orientated single-layer target peptide for the recognition enhancement of CTCs. Covalent and nonvalent binding are to be conducted to mark signal active molecules and nanomaterials on peptides. Signal amplification is implemented with nanomaterials and molecular biotechnology for the improvement of sensitivity. Founded on the above fabrication, the analysis of the abundance and activity of marker protein on the surface of CTCs is to be realized via target peptide-based lab-on-paper.

针对高侵袭恶性循环肿瘤细胞（CTCs）表面间充质样标志蛋白丰度及活性分析检测需求，本项目拟在纸芯片制备中空通道，提高CTCs的分离、富集效率；引入纸电路、流体延时开关及纸超级电容器等纸电子器件，实现光电化学信号的采集、转换与调制；查找结构生物学中提供的高侵袭恶性CTCs上皮-间充质转化标志蛋白，通过文库展示筛选技术筛选与标志蛋白特异性结合位点和构象明确的肽序列；通过数学模型算法、人工改造或组合搭配优化肽功能模体，发展出具有分子识别及信号放大等效果更加突出的靶向肽功能模体；在电极表面修饰亲水性或两亲性物质构建抗污染靶向肽传感界面，界面上组装取向一致单分子层靶向肽，提高细胞的识别能力；通过共价键和非共价键方式对肽标记信号分子、信号纳米材料，研究利用纳米材料和分子生物学技术等制备信号放大结构单元实现信号放大，提高检测方法的灵敏度；实现靶向肽纸芯片高侵袭恶性CTCs表面蛋白丰度及活性分析。

肿瘤标志物的检测对肿瘤早期诊断及术后跟踪有着重要的意义。本项目建立了一系列方法对肿瘤标志物进行了快速检测和肿瘤的即时诊断。通过设计制备功能化微流控纸芯片，设计双极电极纸芯片结构，独立分离检测区域；通过数学模型算法、人工改造或组合搭配优化肽功能模体，筛选出了具有特异性识别的靶向肽功能模体；为了提高检测选择性，通过电极表面修饰亲水性或两亲性物质构建抗污染传感界面；利用纳米材料类酶特性、光电转换性能和标记信号分子实现传感器信号放大，结合CRISPR/Cas、DNA步行者等分子生物学技术，提高检测方法的灵敏度；实现对肿瘤标志物蛋白、核酸的检测。围绕该项目发表高水平论文31篇（SCI 收录）；授权国家发明专利15项；培养博士3名，硕士10名；获山东省高等学校青年创新团队，济南市“高校20条”-科研带头人工作室。