用于质谱流式细胞仪的金属编码聚合物微球的光化学合成研究

Metal-encoded microbeads play an important role in Mass Cytometry immune detection system. In this system, the polymer microbeads must satisfy several important criteria, such as large particle size (>500nm) and high monodispersity, very low variability of metal content from particle to particle, and capable of biological coupling. This project aims to research the synthesis of metal-encoded microbeads by using photoinitiated RAFT dispersion polymerization, and preliminary investigate the applications of the obtained microbeads on Mass Cytometry immune detection system and other similar systems. In the project, we will synthesize body-functional and surface-functional microbeads via photoinitiated RAFT dispersion polymerization by selecting or preparing suitable functional monomers and macro-RAFT agents. For meeting the requirements of Mass Cytometry to the metal-encoded microbeads, we will explore and design new approaches to prepare functional microbeads which is suitable for using as a template to synthesize the metal-encoded microbeads. Then we will modify the microbeads via lanthanide loading and protein coupling by employing the functional groups in/on the microbeads, and finally obtain the lanthanides-encoded microbeads. We will also synthesize the dual-encoded polymer microbeads by introducing both lanthanides and fluorescent dye to the same microbeads, which can be used in both Flow and Mass Cytometry. By further mechanism study and systematic investigation, combined with the existing research foundation, the project will establish a set of effective method of synthesizing metal-encoded microbeads, involving in the high-throughput biological detection fields with our method advantages.

金属编码微球在基于质谱流式细胞仪（Mass Cytometry）的免疫检测系统起关键作用，而该系统对于编码微球有着极高要求，如尺寸（>500nm）、单分散性、金属含量精准控制、生物偶联等。本项目拟研究利用RAFT光分散聚合法合成金属编码微球，并初步探讨所得微球在Mass Cytometry的免疫检测体系及类似系统中的应用。项目将首先探索利用RAFT光分散聚合法合成功能聚合物微球，针对Mass Cytometry对微球的特殊要求考察合成体系和构建新手段，从而得到功能微球载体，并引入镧系元素作为编码。然后利用微球上的功能基团进行蛋白质偶联，最终得到金属负载量可精准控制、可编码量大的镧系编码微球。项目还将尝试合成荧光和金属双编码微球，这种微球可同时在传统流式细胞仪和质谱流式细胞仪中应用。项目最终将建立一套高效的、普适性高的金属编码微球光化学合成方法，以自有技术方法优势介入高通量生物检测高端领域。

本项目拟研究通过 RAFT 光分散聚合法合成金属编码微球，这种金属编码微球可用于质谱流式细胞仪（Mass Cytometry）免疫检测体系。项目立项以来，按计划开展了可用于质谱流式细胞仪的金属编码微球合成研究，包括针对金属负载的微球功能化、功能化大分子RAFT试剂的设计合成、针对生物偶联的微球功能化以及功能微球合成相关机理研究、含镧系元素的编码聚合物微球的合成、双编码微球的合成等。主要成果包括高螯合容量单分散微球的合成方法建立、带有双键的大分子RAFT试剂的合成并用于制备可点击单分散微球、表面富含苯硼酸的聚合物微球的合成、含镧系元素的编码聚合物微球的合成、双编码微球的合成、氧化还原引发RAFT分散聚合法合成单分散微球研究、以纯水为介质合成聚合物微球。总体上，已按计划完成对金属编码微球合成体系的构建。所得成果在单分散功能微球合成方法上向前推进了一大步，对金属编码微球的合成以及在其它领域中应用的微球具有指导作用。但由于一些原因，所得方法未能在质谱流式细胞仪上进行实际试验。相关成果已发表1篇研究论文，并有5项发明专利获授权。