2006 Fiscal Year Final Research Report Summary
Development of High thorough put immunoassay using microchip electrophoresis
| Project/Area Number |
16590464
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Showa University |
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Principal Investigator |
ARAKAWA Hidetoshi Showa University, Pharmaceutical sciences, Professor, 薬学部, 教授 (70129807)
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| Co-Investigator(Kenkyū-buntansha) |
MAEDA Masako Showa University, Pharmaceutical sciences, Emeritus professor, 薬学部, 名誉教授 (00053869)
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| Project Period (FY) |
2004 – 2006
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| Keywords | Capillary electrophoresis / micro chip electrophoresis / immunoassay / electro osmotic flow / DNA labeled antigen / estrogen |
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| Research Abstract |
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
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