环金属铱溶剂配位化合物在电化学发光免疫分析中的应用

ECL immunoassay is known as a highly sensitve and excitation-light-free technology compared with fluorescent immunoassay. Ruthenium(II) tri(bipyridine) complexes [Ru(bpy)32+] are widely-used ECL labeling agents. However, they usually need complicate and tedious labeling procedures and harsh reaction conditions. Furthermore, the emission color of Ru(bpy)32+ is hardly tunable. In recent years, unique ECL properties of cyclometalated iridium complexes have been reported. In our prelimilary studies, we have found that iridium solvento complexes can easily coordinate with histidine residues in protein and the new-formed complexes show obvious ECL signals. Based on this finding, we propose to carry out the research about iridium solvento complexes as antibody labeling agents for ECL immunoassay in this project. These labeling agents can be easily synthesized and have tunable emission color. This project focuses on optimizing the molecular structures of iridium solvento complexes and ECL co-reactants to tune emission color and increase ECL efficiency, and establishing methods of antibody labeling for ECL immunoassay based on iridium solvento complexes.

与荧光免疫分析相比，电化学发光免疫分析具有检测灵敏度高、无激发光源背景干扰等优点。三联吡啶钌金属配合物Ru(bpy)32+是一种常用的电化学发光标记分子，但其发光颜色单一且标记过程繁琐，反应条件苛刻。近年来，环金属铱配合物显示出极具潜力的电化学发光特性。申请人前期研究发现环金属铱溶剂配位化合物可以与蛋白分子表面的组氨酸残基快速配位形成具有电化学发光特性的新化合物。基于此，本项目拟开展环金属铱溶剂配位化合物作为电化学发光免疫分析中抗体标记分子的研究。这类新型的电化学发光标记分子制备方法简单，抗体标记反应条件温和，发光颜色易于通过配体结构进行调节。本项目的研究重点是：从环金属铱溶剂配位化合物主配体的结构设计和电化学发光共反应体系的优化两个方面，调节环金属铱溶剂配位化合物与抗体结合后的发射波长，提高电化学发光效率，为电化学发光免疫分析提供新的抗体标记分子和标记方法。

本项目系统研究了环金属铱配合物分子结构和共反应物类型与电化学发光效率的构效关系，开发了新型的对组氨酸特异性响应的电化学发光探针，设计合成了一系列具有不同主配体结构的新型环金属铱乙腈配合物，用于C-反应蛋白（CRP抗原，临床上一种重要的炎症生物标志物）的电化学发光标记，并进一步建立了基于磁珠分离的CRP抗原电化学发光夹心免疫分析检测新方法，在此基础上并积极开展了发光金属配合物在新型蛋白染色剂、DNA分析检测、光电器件等领域的拓展研究。共计发表SCI 论文9篇（其中SCI I/II区论文5篇），申请或授权发明专利11件。圆满完成了项目计划中的各项研究目标。