量子点-β-内酰胺类药物受体复合物的点击化学制备及其性质研究

The development of simple, quick, and sensitive immunoassay methods is of great importance for the determination of veterinary drug residues in animal origin foods. As compared with the traditional antigen-antibody immunoassay methods, the ligand-receptor systems show a greater performance in broad affinity, and thus have a good prospect in multi-residue analysis. In this study, the penicillin binding proteins (PBPs) will be prokaryotic expressed. According to the previous research, the PBPs and QDs will be functionalized by azide and alkyne, and then the two ligands will be reacted, which made the QDs labeled PBPs. The physical and chemical properties and biological activity of the QDs-PBPs will be characterized by ultraviolet spectrum, fluorescence spectrum, ELISA, and transmission electron microscope techniques. The fluorescence signal will be magnified by Huisgen click chemistry reaction. Based on self-synthesized QDs labeled PBPs, a rapid and sensitive FLISA method will be developed for detection of β-lactam antibiotics in animal foods. In summary, in this work, fluoroimmunoassay based on QDs and PBPs will firstly been evaluated in veterinary drug residue analysis. This study will also confirm that the “click chemistry” can be used in preparation of QDs-based fluorescent markers, which are worthy of in-depth study.

建立简便、快速、灵敏的免疫分析新方法对于有效监控动物源性食品中的兽药残留具有重要的科学意义和应用价值。基于受体-配体反应的配体检测体系较传统免疫分析的抗原-抗体反应在广谱性识别方面正展现出更加优异的性能，因而在多残留分析领域具有很大的发展前景。本项目拟在体外原核表达β-内酰胺类药物的受体-青霉素结合蛋白，并通过对受体蛋白和量子点进行叠氮或炔基化修饰制备量子点-受体复合物。采用紫外光谱、荧光光谱、ELISA测定、透射电镜等分析手段，表征量子点-受体标记物的理化性质和生物活性。在此基础上，利用点击化学的叠氮-炔基Huisgen环加成反应实现量子点荧光信号的大幅扩增，以此为探针，建立动物性食品中β-内酰胺类药物多残留的荧光免疫分析方法。本研究的意义在于将量子点的高灵敏度与受体的广谱性识别相结合用于兽药残留分析领域，为探索兽药残留检测新技术奠定理论基础。

建立简便、快速、灵敏的免疫分析新方法对于有效监控动物源性食品中的兽药残留具有重要的科学意义和应用价值。基于受体-配体反应的配体检测体系较传统免疫分析的抗原-抗体反应在广谱性识别方面正展现出更加优异的性能，因而在多残留分析领域具有很大的发展前景。本项目通过体外原核表达β-内酰胺类药物的受体-青霉素结合蛋白，并通过对受体蛋白和量子点进行叠氮或炔基化修饰制备量子点-受体复合物。采用紫外光谱、荧光光谱、ELISA测定等分析手段，表征量子点-受体标记物的理化性质和生物活性。在此基础上，利用点击化学的叠氮-炔基Huisgen环加成反应实现量子点荧光信号的大幅扩增，以此为探针，建立了动物性食品中β-内酰胺类药物多残留的荧光免疫分析方法。本研究的意义在于将量子点的高灵敏度与受体的广谱性识别相结合用于兽药残留分析领域，为探索兽药残留检测新技术奠定理论基础。