1998 Fiscal Year Final Research Report Summary
Development of soluble TCR and soluble MHC/peptide compelx with high affinity for their ligands
| Project/Area Number |
08557026
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | Kyushu University |
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Principal Investigator |
SASAZUKI Takehiko Medical Institute of Bioregulation, Kyushu University, Professor, 生体防御医学研究所, 教授 (50014121)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUI Yoshinori Medical Institute of Bioregulation, Kyushu University, Associate Professor, 生体防御医学研究所, 助教授 (60243961)
KAMIKAWAJI Nobuhiro Medical Institute of Bioregulation, Kyushu University, Associate Professor, 生体防御医学研究所, 助教授 (90224659)
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| Project Period (FY) |
1996 – 1998
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| Keywords | soluble TCR / soluble MHC / peptide complex / leucine zipper / off rate / affinity |
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| Research Abstract |
We tried in this project to develop soluble MHC class II/peptide complex and soluble TCR with so high affinity as to directly identify and/or isolate their ligands. To facilitate the heterodimer formation, we connected leucine zipper sequence to the extracellular domain of alpha and beta chain of TCR or MHC class II.In addition, the antigenic peptide was covalently bound to MHC class II molecules. In each case, free cysteine was introduced at the C-terminal of one of the leucine zipper sequence, which was used for the target for the specific biotinylation.
Finally, we developed multivalent TCR or MHC class II/peptide complex by coupling the biotinylated molecule with streptavidin. Such multivatent TCR or MHC class II/peptide complex showed remarkably slow off-rate for their ligands in BIAcore analysis and detected the cells expressing the ligand by flow cytometry. Thus, this approach would have wide application for several immunological fields, including T cell development, autoimmunity, transplantation and tumor immunity.
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