| Project/Area Number |
12557239
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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 | 展開研究 |
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| Research Field |
Laboratory medicine
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| Research Institution | National Institute of Advanced Industrial Science and Technology (AIST) (2002)
Tokyo University of Science (2000-2001) |
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Principal Investigator |
FURUKAWA Koji National Institute of Advanced Industrial Science and Technology, Age Dimension Research Center, Senior Research Scientist, 年齢軸生命工学研究センター, 主任研究員 (00297631)
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| Co-Investigator(Kenkyū-buntansha) |
AZUMA Takachika Tokyo University of Science, Research Institute for Biological Sciences, Professor, 生命科学研究所, 教授 (00028234)
NAKAMURA Haruki Osaka University, Institute for Protein Research, Professor, 蛋白質研究所, 教授 (80134485)
KOJIMA Masaharu Kyokuto Seiyaku Kogyo Co.Ltd., Research Development Division, Chief, 研究開発部, 部長代理
ARIKUNI Hisashi Sumisho Biosystems, Kamakura Institute, Director, 鎌倉研究所, 所長
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | antibody / affinity maturation / repertoire / phage display / CDRループ / 抗原結合部位 |
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| Research Abstract |
The purpose of this study is establishment of technical bases for obtaining high affinity antibodies facilitating a performance of protein-chip for clinical investigation as well as the knowledge bases required for the advances of the techniques. To this purpose, the following contributions has been made in this research period.
1)The structural basis for "high evolvability" was clarified using "H3-rules". We found that the evolvability depends on the structural versatility of antigen-combining site, and this could be directly linked to flexibility of the CDR-H3 determined by its base structure and ladder pattern of H-bonds. 2)Thermodynamic property of efficient affinity maturation pathway was examined. We found that the maturation is "zipper -> lock & key" direction. 3)Dynamics shaping antibody repertoire was clarified by an exhaustive repertoire analysis. We found a substantial effect of pre-existing naturally occurring population as well as IgG subclass upon repertoire diversity and the shaping process. 4)In vitro expression and selection system were constructed using a bacteriophage display technique. 5)We found amino acid residues in framework region away from H-L interface correlated well with the pan angle distribution between H- and L-chain of antibodies. This angle reflected potential difference between mouse and human antibodies.
Through these contributions, we are constructing facilitated scheme for obtaining antibodies useful for protein-chip analysis, in which the following basic techniques are involved: immunization, cDNA preparation, antibody expression, in vitro maturation, and fixing antibodies onto protein chips.
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