基于自由流电泳的MRB富集分离新原理与新方法的研究

The research tasks in this project are: (i) the investigation on novel principle and method of MRB created in FFE apparatus; (ii) the study on mechanism of protein mass transfer in the system of FFE-MRB; (iii) the investigation on the theory, method and brief application of MRB-based protein stacking in FFE apparatus; and (iv) the study on the theory, method and use of simultaneous MRB-induced stacking and separation of protein samples in FFE. This project has the following significances: (i) the creation of novel theory of technique of MRB-induced protein separation in FFE, enriching the separation mode of FFE; (ii) the establishment of theory and method of MRB-based protein stacking in FFE, opening new application of FFE in sample condensation of protein in biological field; and (iii) the great improvement of throughput of sample pretreatment in FFE, solving the low throughput of FFE - - - one of the bottleneck issues of FFE.

本课题主要研究内容：（1）、基于自由流电泳（free-flow electrophoresis, FFE）的移动反应界面（moving reaction boundary, MRB）新原理和新方法的研究；（2）、蛋白质在FFE-MRB系统中的传质机理的研究；（3）、基于FFE的MRB高效浓缩模式的理论、方法和初步应用研究；（4）、MRB-介导的同步富集分离新模式理论、方法及其初步应用的研究。意义在于: （1)、建立基于FFE的MRB-介导的蛋白质分离的新原理、新技术和新方法，解决FFE分离模式单一问题；（2)、建立基于FFE的蛋白样品MRB富集新原理和新方法，开拓FFE在生物样品富集等样品前处理方面的应用；（3）、显著提高FFE的样本处理能力，解决FFE样品处理能力低下等的瓶颈性问题。

本课题开展了基于自由流电泳（free-flow electrophoresis, FFE）的移动反应界面（moving reaction boundary, MRB）新原理和新方法的研究，建立了新的MRB模型和理论，阐明了蛋白质在FFE-MRB系统中的传质机理的研究，基本建立了基于FFE的MRB高效浓缩模式的理论、方法和初步应用研究，建立了以线粒体为模型的细胞器富集分离制备新原理、新方法及其相关生物学研究的应用。相关研究已发表较高水平的FFE方面的SCI论文6篇和MRB方面SCI论文8篇。代表性论文如下：.1. iPhone-imaged and Cell-powered Electrophoresis Titration Chip for Alkaline Phosphatase Assay in Serum by MRB, Lab Chip, 2018, 18, 1758-1766..2. A simple chip electrophoresis titration of neutralization boundary with EDTA photocatalysis for distance-based sensing of melamine in dairy product, Anal. Chem., 2018, 90, 6710-6717..3. Double inner standard plot model of electrophoresis titration chip for portable and green protein content assay of milk, Lab Chip, DOI: 10.1039/c8lc01015c..4. Preparation of intact mitochondria using free-flow IEF with post-pH gradient sample injection for morphological, functional and proteomics studies, Anal. Chim. Acta, 2017, 982, 200-208.