Development of High thorough put immunoassay using microchip electrophoresis

本研究采用毛细管电泳和微芯片电泳进行高速免疫分析。首先对毛细管电泳法进行了研究。检出限为10pg，重现性为4.31 （CV%）。其次，研究了稳定电渗透流在微芯片电泳中的产生。这使得应用于免疫测定，并且通过阴离子洗涤剂的涂层方法使由塑料制成的微芯片通道中的壁电离。结果证实了电渗流在良好繁殖中的存在。该方法适用于氨基酸的区带电泳分离。免疫分析法的应用难度较大，灵敏度比微芯片上的毛细管电泳低100倍左右。因此，有必要采用更灵敏的检测系统来开发免疫分析法，然后通过生物和化学发光检测对微芯片电泳进行检测。虽然这是初步的结果，但微M量级的ATP检测成为可能，并提出了通过发光检测进行免疫分析的可能性。此外，本研究还以DNA作为标记抗原的标记物，并考察了嵌入荧光染料的电荷和方法。结果，DNA标记类固醇的分离成为可能，并对其在免疫分析中的应用进行了研究。从本研究的结果来看，毛细管电泳的高速免疫分析成为可能，而微芯片电泳的免疫分析则无法实现。然而，本研究有可能获得许多基本结果。在未来，可以应用于临床领域的高速微芯片免疫分析似乎成为可能，尽管存在抗体，但利用核酸进行适体测定

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