基于金属纳米颗粒@共价键有机框架的C反应蛋白免疫传感器的构建

Covalent Organic Frameworks(COFs) represent an exciting new type of porous and mesoporous organic materials,which are ingeniously constructed with organic building units via strong covalent bonds，appeared after metal organic framework (metal organic framework, MOF). It overcomes the defects of MOF material including skeleton easy to collapse, unstable structure because MOFs are formed by metal ions and organic ligands through coordination bond.COFs possess structure stablity and diversity, controllable pore size etc .Since the seminal work of Yaghi and co-workers in 2005, the rapid development in this research area has attracted intensive interest from researchers with diverse expertise. At present, the cardiovascular disease has become the first killer of human health. Coronary heart disease accounted for more than 50%. C-reactive protein (CRP) is newly found as a marker of inflammation associated with Coronary heart disease. The measurement of serum C-reactive protein (CRP) levels has been given particular interest for early diagnosis and differential diagnosis in clinical cardiovascular diseases. Therefore，there is urgent need for the development of sensitive, accurate, fast detection method for C-reactio protein. On the basis of research on the synthesis and functionalization of 3D COFs, this project plans to synthesis metal nanoparticles @covalent organic framework hybrid materials (COFs) using COFs as template. Furthermore，this hybrid materials will be labeled on antibody of C- reaction protein to construct immunosensor for the accurate and fast detection of C- reaction protein, utilizing the electrochemical signal or absorbance various on the interface caused by specific recognition process between nano biological probe and biological molecules on the sensor interface and c-reactive protein in the sample. This research provides reliable technology platform for the early detection, differential diagnosis and curative effect monitor of diseases such as coronary heart disease (CHD). This study has important significance in clinic for the protection of people's lives and safety.

共价键有机框架（Covalent Organic Frameworks，COFs)是继金属有机框架( MOFs)之后出现的一类新型的微孔和介孔材料，它克服了MOF材料骨架容易发生坍塌的缺点，具有结构稳定多样，孔径可控等特点。目前，心血管疾病已成为人类健康的第一杀手，而冠心病占据了其中50% 以上。C 反应蛋白( CRP)是最新发现的冠心病相关炎性因子，被广泛应用于临床心血管疾病的早期诊断及鉴别。因此，急需发展灵敏、准确，快速的CRP检测方法。本项目拟在深入研究三维COFs的合成及功能化的基础上，实现以COFs为模板合成metal@COFs,并将其作为纳米生物探针标记到CRP抗体上，研制免疫传感器，利用纳米生物探针和传感界面上生物分子与样品中CRP特异性识别过程引起的界面电化学及光度的变化，实现CRP的快速、准确测定，为冠心病的诊治提供技术平台。本研究对于保护人们的生命安全具有重要的临床意义

目前，心血管疾病已成为人类健康的第一杀手，而冠心病占据了其中50% 以上。C 反应蛋白( c-reactive protein, CRP)是最新发现的冠心病相关炎性因子，被广泛应用于临床心血管疾病的早期诊断及鉴别。因此，急需发展灵敏、准确，快速的CRP检测方法。本项目采用溶剂热法通过有机单体合成了四种共价有机框架材料（COFs），其中两种为自主设计合成的新COFs。选用后合成方法将不同的金属颗粒（如Pd、Pt、Au引入到COFs材料上组成复合材料构建电化学C-反应蛋白免疫传感器。代表性工作如下：①首次将COFs材料用于免疫传感器的构建，利用所合成的Pd/COF-LZUl多孔复合材料将CRP抗体（anti-CRP）固定在玻碳电极表面，构建了一种非标记型C-反应蛋白免疫传感器。②提出一种新的免疫传感器检测机理，首次将金属有机框架材料（MOFs）作为信号探针，通过检测MOFs材料中Cu2+的还原峰电流来实现对CRP的检测。③以负载Au颗粒的共价有机框架Au/COF-TpPa-1为固定基质，以氨基化的纳米模拟酶Co3O4为标记物研制了CRP免疫传感器④首次将COFs作为标记物，利用新型共价有机框架（COF-LZU-8）吸附Au颗粒及硫堇制得的复合物（COF-LZU-8/Au/硫堇（Thi）标记CRP抗体，构建了夹心型的CRP生物传感器。⑤利用卟啉具有光电化学（PEC）响应的特性，采用5,10,15,20-四（4-氨基苯基）卟啉作为单体合成一种新的卟啉型共价有机框架（COFs）,用于制备定量检测C反应蛋白的光电适体传感器。⑥以氨基化石墨烯为固定基质将C-反应蛋白抗体固定在电极上，采用中空银铂纳米颗粒标记C-反应蛋白抗体，通过夹心法检测C反应蛋白。该课题研究发表了10篇SCI收录的论文,其中影响因子在5.0以上的3篇;单篇最高影响因子6.32，累计影响因子29.011。另在国内核心刊物上发表了5篇论文，在国内一般期刊上发表论文1篇，获得一项专利授权，超额完成合同上提出的指标。本项目研究对于临床上心血管疾病的诊断具有重要意义。