Development of High thorough put immunoassay using microchip electrophoresis

本研究对毛细管电泳和微芯片电泳快速免疫分析技术进行了研究。首先，使用毛细管电泳的方法进行了检查。方法的检出限为10pg，重复性为4.31（CV%）。接着，检查微芯片电泳中稳定电渗流的产生。这使得能够应用于免疫测定，并且通过阴离子去污剂的涂布方法使由塑料制成的微芯片的通道中的壁离子化。结果，可以确认良好再现中的电渗流。将该方法应用于氨基酸的区带电泳分离。而且，免疫分析的应用比芯片上的毛细管电泳的灵敏度低约100倍更困难。因此，似乎有必要使用更高灵敏度的检测系统来开发免疫分析，接下来，通过生物和化学发光检测来检查微芯片电泳。虽然该结果是初步的，但微M级的ATP检测成为可能，并提出了通过发光检测进行免疫测定的可能性。此外，本研究以DNA作为标记抗原的标记物，并对嵌入荧光染料的电荷和方法进行了考察。结果，标记类固醇的DNA的分离成为可能，并检查了其在免疫测定中的应用。根据本研究的结果，通过毛细管电泳的高速免疫测定成为可能，通过微芯片电泳的免疫测定不能实现。然而，通过这项研究可以获得许多基本结果。在未来，通过使用核酸而不考虑抗体的适体测定，可用于临床领域的高速微芯片免疫测定似乎成为可能

In this study, the high-speed immunoassay using capillary electrophoresis and microchip electrophoresis was examined. First the method using the capillary electrophoresis was examined. The detection limit is 10pg and the reproducibility was 4.31 (CV%). Next, the generation of the steady electroosmotic flow in the microchip electrophoresis was examined. This enabled the application to the immunoassay, and the wall in the channel of the microchip made of the plastic was ionized by the coating method of the anionic detergent. As the result, it was possible to confirm the electroosmotic flow in the good reproduction. This method was applied to zone electrophoresis separation of the amino acid. And, the application of the immunoassay was more difficult than the sensitivity being inferior about 100 times than the capillary electrophoresis on the microchip. Therefore, it seemed to be necessary to use the more supersensitive detection system for development of the immunoassay, and next, it examined microchip electrophoresis by the bio-and chemiluminescence detection. Though as the result, it was preliminary, ATP detection in the micro M order became possible, and the possibility of the immunoassay by the luminescence detection was suggested. In addition, in this study, DNA was used as a marker of labeled antigen, and the charge and method by the intercalation fluorescence dye were also examined. As the result, the separation of the DNA labeled steroid became possible, and the application to the immunoassay was examined. From results of this study, the high-speed immunoassay by the capillary electrophoresis became possible, the immunoassay by the microchip electrophoresis could not be achieved. However, it was possible to obtain many basic results by this study. In the future, high speed microchip immunoassay which can be used in the clinical field seems to become possible by using aptamer assay using the nucleic acid in spite of antibody