Study On Wide-bore Electrophoresis And One-step Electrophoretically Mediated Microanalysis Technique

With the rapid development of science and technology, the trend for in-depth research on object compounds has been finality in the post-genome era. Capillary electrophoresis (CE), which played an important role in Human Genome Project, exposed its limitations in dealing with large volume of samples. Therefore, the tiny sample loading capacity becomes the major obstacle to extending the application scope of CE.In order to increase the sample loading capacity while maintaining the high separation efficiency in column electrophoresis, a new system called wide-bore electrophoresis (WE) was adopted in the thesis based on the principle of shell and tube heat exchanger. The Joule heat generated during the electrophoresis process could be removed in situ in real time thoroughly by the inner cooling water. The experimental data indicated that the heat sink was effective and could make the function of the applied potential and current in the WE system matches Ohm plots well. Some acidic or alkaline model compounds were selected to evaluate the separation efficiency of the novel system and a separation efficiency as high as over 60, 000 plates/m was achieved for some compounds. The biggest advantage of this WE system is its potential as a preparative or semi-preparative electrophoretic separation technique due to the maximum sample loading capacity of 1.0μL or more.The larger sample loading capacity entitled the WE system for multi-dimensional operations. Thus, the delivery properties of object compounds were tested by using ponceau 4R as the model. The feasibility of the compound's delivery was validated by the experiment.In future work, one intention of establishing WE system is to develop a multi-dimensional high-throughput medical screening platform. Because enzyme analysis is a vital unit in this medical screening, a one-step enzyme microanalysis technique was developed on the basis of Electrophoretically Mediated Microanalysis (EMMA) method. Furthermore, the enzyme kinetics constant of alkaline phosphatase (ALP) was determined using the traditional method and the one-step method. The experimental results demonstrated that the one-step determination method for enzyme kinetics constant was accurate and sample saving. Therefore, the one-step enzyme analysis method proposed in this thesis combined with WE system to perform multi-dimensional screening is worth of in-depth research and further development.