| Project/Area Number |
15K08629
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Kobe Pharmaceutical University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
大山 浩之 神戸薬科大学, 薬学部, 助教 (80572966)
森田 いずみ 神戸薬科大学, 薬学部, 助手 (20299085)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 抗体 / 一本鎖Fvフラグメント / 変異体 / ライブラリー / 選択方法 / パンニング / 変異抗体 / バイオマーカー / 進化分子工学 / ファージ提示 / マイクロアレイ / デジタル検出 |
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| Outline of Final Research Achievements |
Phage-display-based molecular evolution potentially provides antibody mutants with greater diagnostic utilities than conventional antibodies. The panning, in which whole members were reacted simultaneously to immobilized antigens, has still been used to isolate evolved mutants in diverse antibody libraries, but often missed valuable species. Thus, we developed a new selection strategy to solve this crucial problem. Bacterial clones in a library, each harboring a single-chain Fv fragment (scFv) gene, were cultured separately in microwells, and scFv-displaying phage generated therein was examined for their antigen-binding property using a bioluminescent assay. One-shot application for an scFv library against cortisol provided several scFv clones with improved affinities, none of which could be isolated through a repeated trials of conventional panning. Thus we evaluated that the present method should be of significant utility for generating high-performance diagnostic antibodies.
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