| Project/Area Number |
14572039
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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 |
Physical pharmacy
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| Research Institution | Nihon University |
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Principal Investigator |
SHINOMIYA Kazufusa Nihon University, College of Pharmacy, Associate professor, 薬学部, 助教授 (70215995)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | countercurrent chromatoeraphy / aqueous two-phase partitioning / protein separation / cell separation / chromatosgraphy / separation analysis / biological macromolecule / blood cell / 水生二層分配法 / 生態高分子 |
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| Research Abstract |
1. Protein separation using the nonsynchronous coil planet centrifuge (CPC)
The nonsynchronous CPC was employed to investigate the optimal separating conditions and the effect of planetary motion of coiled column on the partition efficiency using the protein separation with aqueous-aqueous polymer phase systems. The separation was examined using three kinds coiled column with each different volume (11, 24 and 39 mL), a two-phase system composed of 12.5% (w/w) polyethylene glycol 1000 and 12.5% (w/w) dibasic potassium phosphate and a set of stable proteins for samples including cytochrome C, myoglobin and lysozyme. The best separation was obtained using the 39 mL multilayer coil at the head to tail elution mode with the revolution at 800 rpm and the rotation at 10 rpm, where the similar result was obtained in the eccentric coil under same experimental conditions. The further experiments also revealed that the best result was obtained in the head to tail elution mode by the clockwise (CW) coil rotation for the lower mobile phase or by the counterclockwise (CCW) coil rotation for the upper mobile phase, both under CCW revolution of the rotor.
2. Elutriation of blood cell components by the nonsynchronous CPC
The nonsynchronous CPC was applied to the separation of blood cell components using a physiological buffer solution. The optimal condition for the separation of sheep blood samples was performed at the CCW revolution and the CW rotation of coiled column by the head to tail elution mode. Under the high speed of revolution of 600-1000 rpm, sheep and human blood samples were each completely separated and eluted into two peaks, where all cell components other than erythrocytes such as platelets and leukocytes were eluted at 0 rpm and erytbrocytes at 10 rpm of coil rotation. The normal elution pattern was achieved at 800 rpm of revolution for sheep blood and at 700 rpm for human blood samples.
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