基于纳米材料自组装和环境敏感型自供能微流控-SERS技术的肺癌早期检测新方法研究

Due to the high morbidity and mortality of lung cancer in China, development of efficient methods for early diagnosis is extremely significant. The use of tumor markers as prognostic and prediction in patients with lung cancer is one of the effective approaches. Based on the characteristics of tumor markers, such as trace concentration, complicated components of the sample, as well as the requirement of combined detection of multiple tumor markers, we will explore and develop a new method based on the self-assembly of nanomaterials, stimuli-responsive self-actuated microfluidic devices, and surface-enhanced Raman spectroscopy (SERS) detection. First, a large-area, sensitive, highly uniform, and controllable SERS substrate with three-dimensional (3D) nanostructures will be fabricated via the self-assembly of nanomaterials inside microfluidic channels. Then, the capture ligands which have specific selectivity for tumor markers are immobilized on the plasmonic nanostructured pattern via directional coupling or ordered arrangement. These ligands will include antibody, aptamer, and molecularly imprinted polymers. A self-powered integrated microfluidic device without external valving or pumping mechanisms is designed by regulating the stimuli-responsive elements from nanomaterials which are capable of driving flow and actuating the valve. The aim of this study is to design a nanobiosensor enabling efficient enrichment and detection of multiple tumor markers from blood serums of lung cancer patients, which exhibit high selectivity, accuracy and sensitivity. Investigation of this study can provide a useful tool for early diagnosis of lung cancer.

肺癌是我国发病率和死亡率最高的一种癌症，实现肺癌的早期检测对降低肺癌死亡率具有重要的研究意义，而肿瘤标志物检测是实现肺癌早期诊断的重要手段之一。针对肺癌早期患者临床样品成分复杂、肿瘤标志物浓度低、需要联合检测等特点，本项目将研究和开发基于纳米材料自组装、环境敏感型自供能微流控芯片和表面增强拉曼光谱（SERS）技术的肺癌早期检测新方法。首先，功能纳米材料在微流控芯片的微通道内自组装构建大面积、均匀可控、高灵敏度的三维纳米结构SERS基底；其次，具有特异选择性的靶向配体（抗体、核酸适配体）和分子印迹聚合物在微界面上发生定向偶联和有序组装；通过环境敏感型纳米材料对外界刺激的响应性实现微流控芯片上液流的自动驱动和控制；最终通过微流控-SERS技术实现多种肿瘤标志物的高通量富集和超高灵敏度检测，为肺癌等重大疾病的早期诊断和筛查提供理论和实践基础。

肺癌是我国发病率和死亡率最高的一种癌症，实现肺癌的早期检测对降低肺癌死亡率具有重要的研究意义，而肿瘤标志物检测是实现肺癌早期诊断的重要手段之一。针对肺癌早期患者临床样品成分复杂、肿瘤标志物浓度低、需要联合检测等特点，本项目研究和开发了基于纳米材料自组装、环境敏感型自供能微流控芯片和表面增强拉曼光谱（SERS）技术的肺癌早期检测新方法。取得了如下的研究成果：（1）通过纳米材料自组装，设计合成了分子印迹纳米生物探针、核酸适配体纳米生物探针、金属有机框架材料生物探针，结合SERS检测技术，实现了肿瘤标志物的高选择性检测和成像，检测限达10-14 mol/L。（2）通过化学定向进化新方法设计合成了非生物纳米人工抗体，对肿瘤标志物的亲和力常数KD低达10-11 M，与抗体的相当，并具有良好的选择性。（3）设计合成了温度敏感性分子印迹整体柱聚合物，成功实现了对蛋白质标志物的选择性捕获和释放；基于纳米金整体柱聚合物板构建了高灵敏度拉曼增强基底，拉曼增强因子为1.3×105。（4）设计合成了自供能微流控芯片结合核酸适配体-SERS纳米生物探针，实现了血清样品中CEA、NSE、ProGRP6、microRNA21、microRNA205、microRNA210等6种肺癌标志物的高通量富集和超高灵敏度检测，为肺癌等重大疾病的早期诊断和筛查提供理论和实践基础。项目资助相关研究成果发表SCI收录论文19篇，申请国家发明专利8项。项目培养博士研究生3名，培养硕士研究生5名，参加16次国际国内会议介绍相关成果。项目投入经费65万元，支出50.7007万元，各项目支出基本与预算相符，剩余经费14.2993万元，剩余经费计划用于本项目研究后续支出。