基于电纺纤维固相传感平台的肿瘤标志物可视化分析方法与技术

Highly sensitive and selective quantification of tumor biomarkers is very significant for early disease screening, diagnosis and treatment and drug research. Wherein, solid sensing platform play a vital role in point-of-care test (POCT). This project aim to exist some deficiencies: low sensitivity, poorly visual effect, unsatisfactory reproducibility in some conventional methods such as paper-based strips. To further address molecular-recognized mechanism on solid sensing interfaces, this project use flexible electrospun fibers as substrates to construct the solid sensing platform for visual detection toward tumor biomarkers. At first, using optical probes including gold nanorods (Au NRs) or nanobipyramids (Au NBPs) and luminescent quantum dots (QDs) or carbon dots (CDs) with strong luminescence as signal output elements, aptamer and antibody as target-discriminated sites, we decorate the two key parts on electrospun fibrous surfaces by chemical bonds or affinity labeling to construct the visually sensing interfaces. Second, we introduce some signal amplified strategies such as catalytic hairpin assembly, hybridization chain reaction or metal ions-mediated amplification to improve the sensitivity of solid fibrous sensing platform, which can be further employed to visually quantify the tumor biomarkers in complex physiological samples. Third, we study the color molecule-recognized mechanism and conversion mechanism on solid electrospun fibrous sensing interfaces, and try to establish the relationship between the micro-structures of electrospun fibers and its analytical performances. Finally, the expected outcomes of this project will provide some valuable theoretical evidence and technique support for the early screening, diagnosis and treatment of tumor.

高灵敏、高选择地检测肿瘤标志物对癌症早期筛查、诊疗和药物研究有着重要意义，其中，固相传感平台在即时检测方面更是扮演着重要角色。本项目针对传统试纸等固相传感方法灵敏度低、可视化效果不理想、重现性差等问题，以解决固相传感界面分子识别机制问题为目标，构建柔性电纺纤维固相传感平台用于肿瘤标志物的可视化分析：(1)以金纳米棒/双锥、量子点或碳点等纳米光谱探针为信号输出元件，适配体或抗体为靶物识别位点，通过化学键、生物嫁接等方式将其修饰到纤维表面，构建可视化传感界面；(2)引入发卡自组装催化、杂交链式反应、金属离子介导等信号放大策略提高其对靶物检测的灵敏度，并用于实际复杂生理样品中标志物的可视化分析；(3)研究固相膜传感界面发生的分子识别机制及变色机制，通过调控电纺纤维的光学信号，建立电纺纤维膜结构特性与检测性能之间的构效关系。本项目预期成果有望为肿瘤早期筛查、诊疗提供非常有价值的理论依据和技术支持。

高灵敏、高选择地检测肿瘤标志物对癌症早期筛查、诊疗和药物研究有着重要意义，其中，固相传感平台在即时检测（point-of-care testing, POCT）方面更是扮演这重要角色。本项目针对传统试纸等固相传感方法灵敏度低、可视化色度效果单一、重现性差等问题，已解决固相传感界面分子识别机制问题为目标导向，构建了多种基于基纳米结构或者荧光颗粒的柔性电纺纤维固相传感平台用于肿瘤标志物的可视化分析。主要的研究内容有：（1）目标物诱导的三足G4-DNAzyme催化氧化腐蚀金纳米棒产生多色电纺膜可视化效果，用于开发生物标志物的POCT技术。（2）生物酶催化诱导FeOOH分解产生大量的Fe2+，Fe2+作为芬顿试剂，诱导金纳米双锥腐蚀，导致金纳米双锥纵向LSPR吸收峰蓝移产生色度丰富的电纺膜颜色，用于乙酰胆碱酯酶及其抑制剂药物的筛选。（3）基于ꞵ-半乳糖苷酶（ꞵ-Gal）催化底物触发CoOOH分解，导致Co2+和I−之间的竞争作用，借助金纳米双锥作为可视化光谱探针，用于ꞵ-Gal及微生物细菌的可视化分析。（4）受到上述研究内容中涉及到的信号放大策略，借助纳米酶高强度的催化特性，还构建了一种基于Fe凝胶的适体传感器用于食品中伏马毒素的可视化分析。（5）紧密围绕本项目分子识别机制问题的探讨，本项目基于DNA纳米机器作为信号放大策略，设计了熵驱动、酶驱动、双步行DNA纳米机器用于前列腺癌特异抗原（PSA）、黏膜蛋白（MUC1）、microRNA的高灵敏、高选择性检测。（6）基于上述研究内容，还进一步探讨了电纺纤维膜其传感界面发生的分子识别机制问题、变色机理，重要的是对于传感膜稳定显色的储存模式有了新的认识。本项目的研究成果为肿瘤早期筛查、诊疗、药物研究将提供非常有价值的理论依据和技术支持。本项目共培养已毕业硕士研究生1名，研究发表SCI论文6篇，其中，中科院分区一区论文5篇，四区（中文核心）论文1篇，累积影响因子：51.334，申请并授权中国发明专利1项。