基于芳基重氮盐修饰技术的细胞因子电化学免疫生物传感器的构建

Cytokine detection is an important index to judge the immune function of organism, and is important to the early diagnosis of disease, the judgment of curative effect and the monitoring of cytokine therapy. The ultralow concentration of cytokines in serum presents the challengings for the accurate and fast cytokine detection. Based on the principle of aryldaizonium salt reductive adsorption, this project plans to modify the composite nanomaterials of reductive graphene oxides and gold nanoparticles (AuNP), mixed aryldiazonium salts between zwitterionic molecules and cytokine capture antibodies to form a sensing interface which can concentrate the specified cytokines. After that, the sensing interface will interact with the AuNPs modified with cytokine detection antibodies and redox probes to develop a sandwich immunosensing system for detection of localized cytokines. The main target investigations include: the performance of nanomaterials on the preconcentration of cytokines on sensing interface, the performance of aryldiazonium salt of zwitterionic molecules on resisting the non-specific protein adsorption, the influence of AuNPs on signal amplifying, and the performance of the whole immunosensing system for detection of localized cytokines in serum. Based on the studies in this project, we wish we can step further on sensitivity, stability, and selectivity for cytokine detection by electrochemical immunosensors. In addition, we wish this project can provide guidance on the quick point-of-care diagnosis of cytokines.

细胞因子的检测是判断机体免疫功能的一个重要指标，对于疾病的早期诊断、疗效判断及细胞因子治疗监测等都有重要意义。细胞因子在人体血液中的超低含量对细胞因子的准确快速检测提出了挑战。本项目根据芳基重单盐还原吸附的原理，在电极表面修饰上还原石墨烯和金纳米颗粒（AuNP）复合纳米材料、混合芳基重单盐两性分子和细胞因子捕捉抗体，形成一种可富集特定细胞因子的传感器表面。然后，将修饰有检测抗体和氧化还原分子的AuNP作用于次传感器表面，构建一种夹心型电化学免疫生物传感器，用于局部细胞因子的检测。本项目深入研究复合纳米材料对细胞因子预富集的表现、两性分子在血液样品检测中抑制非特异性蛋白质吸附上的表现、修饰有检测抗体的AuNP对检测信号放大的影响、及所构建的细胞因子免疫生物传感器的工作性能。通过本项目的研究希望能提高细胞因子电化学免疫分析法的灵敏性、稳定性和选择性，并对细胞因子的快速临床诊断能提供指导意义。

细胞因子的检测是判断机体免疫功能的一个重要指标，对于疾病的早期诊断、疗效判断及细胞因子治疗监测等都有重要意义。细胞因子在人体血液中的超低含量对细胞因子的准确快速检测提出了挑战。本项目根据芳基重单盐还原吸附的原理，在电极表面修饰上还原石墨烯和金纳米颗粒（AuNP）复合纳米材料、混合芳基重单盐两性分子和细胞因子捕捉抗体，形成一种可富集特定细胞因子的传感器表面。然后，将修饰有检测抗体和氧化还原分子的AuNP作用于次传感器表面，构建一种夹心型电化学免疫生物传感器，用于局部细胞因子的检测。本项目深入研究了复合纳米材料对细胞因子预富集的表现、两性分子在血液样品检测中抑制非特异性蛋白质吸附上的表现、修饰有检测抗体的AuNP对检测信号放大的影响、及所构建的细胞因子免疫生物传感器的工作性能。本项目的研究明显地提高了细胞因子电化学免疫分析法的灵敏性、稳定性和特异性。并对细胞因子的快速临床诊断能提供了指导意义。同时，本项目超计划完成任务，除了构建了一系列的细胞因子免疫生物传感器，还开发了有巨大应用前景的基于适配体的实现检测细胞因子的体系，和细胞因子激活的药物释放体系，实现了用于体内免疫系统精准诊疗一体化装置。共发表文章37篇，专利5个。