| Project/Area Number |
15350041
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
OTSUKA Koji KYOTO UNIVERSITY, GRADUATE SCHOOL OF ENGINEERING, PROFESSOR, 工学研究科, 教授 (70183762)
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| Co-Investigator(Kenkyū-buntansha) |
HAGINAKA Jun MUKOGAWA WOMEN'S UNIVERSITY, SCHOOL OF PHARMACEUTICAL SCIENCES, PROFESSOR, 薬学部, 教授 (20164759)
KITAGAWA Fumihiko KYOTO UNIVERSITY, GRADUATE SCHOOL OF ENGINEERING, ASSISTANT PROFESSOR, 工学研究科, 助手 (20362452)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,600,000 (Direct Cost: ¥14,600,000)
Fiscal Year 2004: ¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 2003: ¥9,000,000 (Direct Cost: ¥9,000,000)
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| Keywords | protein / capillary electrophoresis(CE) / microchip electrophoresis / mass spectrometry(MS) / isoelectric focusing / chiral separation / CE-MS / thermal lens microscopy / MCE-MS |
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| Research Abstract |
Toward the high performance separation and detection of proteins by capillary electrophoresis(CE) and microchip electrophoresis(MCE), following five subjects were mainly investigated.
1)Development of a high performance analysis method by microchip isoelectric focusing(MCIEF). Four model proteins were successfully separated within 210 s by MCIEF, where good linear relationship between a pI value of the protein and its focused position.
2)Development of a highly efficient chiral separation technique by capillary electrochromatography and microchip electrochromatography using immobilized proteins. Avidin was used as a chiral selector and four pairs of enantiomers were successfully resolved.
3)Improvement of detection sensitivity in MCE using on-line sample preconcentration. Under a micellar electrokinetic chromatography (MEKC) mode in MCE, successful on-line sample preconcentration by SRW was achieved by using a newly designed and fabricated T -cross type channel quartz chip.
4)Improvement of detectability in CE and MCE by using thermal lens microscopy (TLM). In a microchip MEKC-TLM system with sweeping as an on-line sample preconcentration technique, the limit of detection of 4 nM or 8 amol in absolute amount injected for Sunset Yellow was obtained.
5)Development of a high performance MCE system with mass spectrometric (MS) detection. An MCE-MS system with an electrospray ionization interface was developed, where some peptides and proteins were successfully separated and detected.
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