| Project/Area Number |
15360442
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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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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Kobe University |
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Principal Investigator |
KATOH Shigeo Kobe University, Graduate School of Science and Technology, Professor, 自然科学研究科, 教授 (20026272)
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| Co-Investigator(Kenkyū-buntansha) |
KATUSDA Tomohisa Kobe University, Dept.Chemical Science and Engineering, Assistant Prof., 工学部, 助手 (50335460)
SHIOMI Naofumi Kobe College, Dept.Human Science, Professor, 人間科学部, 教授 (20299077)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2004: ¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2003: ¥9,600,000 (Direct Cost: ¥9,600,000)
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| Keywords | Colony lift assay / Antibody-coupled liposome / Micro-assay / Antibody library / Production of antibody by secretion / Biopanning / マイクロブロットアッセイ / バイオパンニング / 一本鎖抗体生産 / 疎水性膜 |
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| Research Abstract |
In the fields of development of pharmaceuticals, medical diagnosis-analysis and bioprocesses, production of variety of antibodies, detection of the interactions between antibody and protein, screening methods of antibodies and effective production processes of antibodies are essential. In this work, we developed the simple and rapid methods to obtain antibody ligands for recognition and detection of target proteins, based on our recent researches on production of large libraries of antibodies, a high sensitivity detection method of proteins and effective production of antibodies by secretion from yeast cells.
1.A colony lift assay for detection of antibody secreted from E. coli was developed by use of antigen-adsorbed PVDF membrane and liposome immunoblotting assay. By use of this method, high density and micro-assay was possible and the number of samples treated was increased.
2.By use of HRP encapsulating immunoliposomes and hydrophobic membranes, an immunoblotting assay method with two orders of magnitude higher sensitivity, compared with the conventional immunoblotting assay using HRP-conjugated antibody, was developed.
3.A large library of whole antibodies was obtained based on mating of yeast cells transformed by libraries of H- and L-chains, and aproduction method of antibodies by secretion from yeast cells was established.
4.An affinity biopanning method utilizing immunoliposomes was developed and it was applied to biopanning selection with a high efficiency.
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