| Project/Area Number |
10145102
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas (A)
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| Allocation Type | Single-year Grants |
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
MATSUNAGA Tadashi DEPT.BIOTECHNOLOGY., TOKYO UNIV.AGRICULTURE & TECHNOLOGY, PROF., 工学部, 教授 (10134834)
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| Co-Investigator(Kenkyū-buntansha) |
KOBATAKE Eiri SCHOOL OF BIOSCIENCE AND BIOTECHNOL., TOKYO INSTITUTE OF TECHNOL., ASSOCIATE PROF., 生命理工学部, 助教授 (00225484)
GOTO Masahiro DEPT.CHEMICAL SYSTEM ENGINEERING, GRADUATE SCHOOL OF ENGINEERING, KYUSHU UNIV., ASSOCIATE PROF., 工学部, 助教授 (10211921)
KOMIYAMA Makoto RESEARCH CENTER FOR ADVANCED SCIENCE AND TECHNOLOGY, THE UNIV.TOKYO, PROF., 大学院・工学系研究科, 教授 (50133096)
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| Project Period (FY) |
1998 – 2001
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| Keywords | molecular architecture / bacterial magnetic particle / molecular imprinting / artificial receptor / bio-targeting / surfactant-enzyme complex / molecular recognizing protein / immunoliposome |
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| Research Abstract |
l) We have successfully constructed molecular architectures on bacterial magnetic particles (BMPs) using MagA or Mms16 as an anchor protein by two different approaches. First, target protein fused with the anchor protein MagA was directly inserted into the BMP membrane in vitro using sonication. Second, magA or mms16-target hybrid gene was constructed and the fusion protein was displayed on the BMP surface in vivo. Furthermore, The molecular architectures were applied to detection system for several biomarkers for diagnosis of diabetes and estrogen receptor binding assay.
2) Our purpose is to create artificial receptors which can recognize nano-scaled large molecules in aqueous media as natural receptors do. The strategy we have proposed for the purpose is to build up several host molecules so that each of them fits the designated portion of the target large guest. We chose cyclodextrins (CyDs) as a functional monomer since they bind angstrom-sized guests through apolar interaction in p
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rotic media. CyDs were successfully crosslinked with diisocyanates in the presence of various template molecules in DMSO or water in which CyDs can form complex.
3) We have investigated the bio-targeting effect of liposomes that includes the newly synthesized sugar lipids. The degree of targeting effect of the sugar included liposome was evaluated by the binding test with a specific lectine that possesses a corresponding recognition site on the protein surface. The sugar lipid exhibited an excellent bio-targeting effect.
4) IgG-binding protein was genetically expressed and lipid-modified in a site-directed manner in Escherichia coli.-The lipid-modified protein was translocated at the bacterial membrane, and it was purified with IgG-Sepharose by affinity chromatography. The resulting protein, which retained IgG-binding activity, was displayed on the surface of liposome by anchoring the lipid-tail into the hydrophobic layer on the liposome membrane. Immunoliposomes were prepared by introducing antibody molecules onto the surface of proteoliposomes with highly oriented manner. Finally, they were applied to a novel immunoassay system. Less
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