Advanced phage-display method with high efficiency by magnetic orientation of rod-like viruses
| Project/Area Number |
20H02791
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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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| Review Section |
Basic Section 35010:Polymer chemistry-related
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Ishida Yasuhiro 国立研究開発法人理化学研究所, 創発物性科学研究センター, チームリーダー (20343113)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2022: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2021: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2020: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
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| Keywords | ファージディスプレイ法 / 分子進化工学 / 親和性選択 / ライブラリ / 磁場配向 / ファージディスプレイ 法 |
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| Outline of Research at the Start |
標的に対し強固に結合するペプチドを得る上で、ファージディスプレイ法は有用な手法であるが、提示ペプチドよりはるかに巨大なファージ外殻が標的に非特異的に吸着し、選別効率を低下させることが長年の問題であった。本研究では、平滑な基板に固定された標的物質にファージライブラリを相互作用させる際、磁場でファージを垂直方向に配向することにより非特異吸着を軽減するという、新たなファージディスプレ法の開拓を目指す。
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| Outline of Final Research Achievements |
Although phage display selection using a library of M13 bacteriophage has become a powerful tool for finding peptides that bind to target materials on demand, a remaining concern of this method is the interference by the M13 main body, which is a huge filament >1000 times larger than the displayed peptide, and therefore would nonspecifically adhere to the target or sterically inhibit the binding of the displayed peptide. Meanwhile, filamentous phages are known to be orientable by an external magnetic field. This work proved that the magnetic orientation of M13 filaments vertical to the target surface significantly affects the selection. When the target surface was affinitive to the M13 main body, this orientation notably suppressed the nonspecific adhesion. Furthermore, when the target surface was less affinitive to the M13 main body and intrinsically free from the nonspecific adhesion, this orientation drastically changed the population of M13 clones selected from the library.
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| Academic Significance and Societal Importance of the Research Achievements |
2018年ノーベル化学賞の対象ともなったファージディスプレイ法が、その創設以来抱える問題（巨大なファージ外殻による吸着選択効率の低下）に対し、化学物質の添加を必要とせずにクリーンな物理的刺激のみを用い、様々な標的物質に対し適応できる解決法を提示した。
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Report
(4 results)
Research Products
(17 results)
