| Project/Area Number |
16613008
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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 |
食の安全
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| Research Institution | Kagoshima University |
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Principal Investigator |
HASHIGUCHI Shuhei Kagoshima University, Faculty of Engineering, Research Associate, 工学部, 助手 (40295275)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIMURA Kazuhisa Kagoshima University, Faculty of Engineering, Professor, 工学部, 教授 (80127240)
ITO Yuji Kagoshima University, Faculty of Engineering, Assistant Professor, 工学部, 助教授 (60223195)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Phage display / Human antibody / Prion / Antibody engineering / Conformation / Food / Diagnosis / Neurodegenerative disorders / 抗体エンジリアリング / 脳神経疾患 |
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| Research Abstract |
Recombinant prion protein (PrP : aa23-231) was refolded into soluble from rich in β-structure at pH 4.0, as determined by circular dichroism (CD). The soluble β form (β-PrP) exhibited partial resistance to proteinase K digestion, and composed of small spherical particles (3 nm in diameter estimated by atomic force microscopy). Direct panning was done against β-PrP using a human scFv (single chain Fv)-displaying phage library. After two rounds of panning, we isolated two clones (β-PrP7 and β-PrP30) specifict to β-PrP. We tested the binding activity of these clones against β-PrP preparations that had been incubated at 4℃ for 0, 4 or 14 days. The binding activity of both clones became stronger depending on the incubation time. These results indicate that the content of β-PrP conformers recognized by β-PrP7 or β-PrP30 has increased dynamically in the solution at 4℃, which process is detectable by these particular conformation-specific scFv clones.
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