| Project/Area Number |
09557030
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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 |
Immunology
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| Research Institution | Fujit Health University |
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Principal Investigator |
KUROSAWA Yoshikazu Institute for Comprehensive Medical Science Professor, 総合医科学研究所, 教授 (10109259)
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| Co-Investigator(Kenkyū-buntansha) |
IBA Yoshitaka Institute for Comprehensive Medical Science Research Associate, 総合医科学研究所, 助手 (90298547)
AKAHORI Yasuaki Institute for Comprehensive Medical Science Researchi Associate, 総合医科学研究所, 助手 (80221711)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥10,800,000 (Direct Cost: ¥10,800,000)
Fiscal Year 1999: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1997: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | recombinant DNA / antigen antibody interaactions / CDR framework region / antibody library / phage-display system / ファージ抗体 / 抗体レパートリー / ヒト抗体 / 抗原抗体反応 / error-prone PCR / 抗体-抗体反応 / 人工抗体 / ステロイド抗原 / チトクロームC抗原 / γ-グロブリン製剤 / error-pronePCR / ヒト型抗体 |
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| Research Abstract |
We examined the conditions for constructing libraries of antibodies from which antibodies can be isolating only by screening with various antigens. It has been well known that antibodies bind specifically to antigens through the structures formed by six CRDs (complementarity-determining regions) that are located in V regions of H and L chains. Our basic design for construction of libraries of antibodies is diversification of amino acid sequences of six CDRs. We used the phage -display system. First, we compared the form of antibody expressed on the phage surface among Fab form, single chain Fv form and Fv form and we adopted the Fab form of antibody. Next, we have developed the strategy for changing antigen specificity using steroid antigens and antibodies. Thirdly, we performed CDR-grafting experiments and found that amino acids at several positions in the framework regions and their combinations gave substantial effects on the structures formed by the CDRs. Thus, we obtained various precious informations upon antigen-antibody interactions and the characteristics of phage-display system. For construction of libraries of antibodies. however, we adopted completely different strategies later on.
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