| Project/Area Number |
09640731
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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 |
分離・精製・検出法
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| Research Institution | Ehime University |
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Principal Investigator |
MANABE Takashi Ehime University, Faculty of Science, Department of Chemistry, Professor, 理学部, 教授 (60094281)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMAZAKI Yoji Ehime University, Fculty of Science, Assistant, 理学部, 助手 (80284389)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | plasma proteins / capillary electrophoresis / myeloma / isoelectric focusing / SDS electrophoresis / high-sensitivity analysis / high-resolution anaiysis / two-demensional gel electrophoresis / 免疫グロブリン / タンパク質 / 血漿 / 血清 |
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| Research Abstract |
We aimed to develop techniques to analyze human plasma mycloma proteins in high sensitivity and high resolution. Microscale two-dimensional gel clectrophoresis and capillary electrophoresis were the candidates, but we found that two-dimensional gel clectrophoresis requires many steps of manual handling to obtain the separation patterns and additional processes of image analysis arc necessary to quantitate each separated proteins. Then we examined two separation principles of capillary electrophoresis, capillary isoelectric focusing and capillary SDS elcetrophoresis, for the analysis of mycloma proteins. Electrophoretic conditions for high-resolution and high-sensitivity separation of human plasma proteins have been developed for the two separation principles of capillary electrophoresis. Employing the established separation conditions, normal human plasma proteins have been separated as about 60 peaks/shoulders within 60 minutes by capillary isoelectric focusing and as about 30 peaks/shoulders within 70 minutes by capillary SDS electrophoresis. Major plasma proteins and mycloma proteins can be identified and quantitated on-column during separation by their UV absorbance. Our results show that capillary isoelectric focusing and capillary SDS electrophoresis are quite effective for high performance separation and analysis of human plasma mycloma proteins, when the separation conditions we have established were employed.
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