Development of tecnologies for production of high affinity human antibodies and its application to specific therapies and risk evaluation.
| Project/Area Number |
14360210
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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 |
Applied molecular and cellular biology
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
SHIRAHATA Sanetaka Kyushu University, Faculty of Agriculture, Professor, 大学院・農学研究院, 教授 (90154377)
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| Co-Investigator(Kenkyū-buntansha) |
KATAKURA Yoshinori Kyushu University, Faculty of Agriculture, Associate Professor, 大学院・農学研究院, 助教授 (50264106)
TERUYA Kiichiro Kyushu University, Faculty of Agriculture, Research Associate, 大学院・農学研究院, 助手 (10273971)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥15,100,000 (Direct Cost: ¥15,100,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2002: ¥10,800,000 (Direct Cost: ¥10,800,000)
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| Keywords | in vitro immunization / human antibody / adjuvant / phaze display method / allergen / TNF-α / antibody for peptides / large-scale production / ファージディスプレー法 / コメアレルゲン / アレルギー / IgE |
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| Research Abstract |
(1)Clarification of the mechanisms of in vitro immunization : In in vitro immunization, it was indispensable to remove cells suppressing antibody production by treatment of peripheral blood lymphocytes with leucyl methyl ester (LLME). Analysis revealed that target cells of LLME was immature dendritic cells, enabling to develop a new method to efficiently induce in vitro immunization by removing immature dedritic cells, instead of using LLME (patent application). Furthermore, instead of muramyl dipeptide as an adjuvant, CpG oligonudleotides were used to induce B cells secreting antigen-specific antibodies efficiently and reproducibly.
(2)Efficient acquisition of variable region of human antibody genes by usinga hybrid system of in vitro immunization and phage display method : Lymphocytes induced antigen-specific antibody production by in vitro immunization were used for phage display method to obtain variable region of antigen- specific human antibody gene efficiently and reproducibly.
(3)Production of human antibodies for peptides. : In order to enhance the wide use of in vitro immunization method, antibodies far peptides was produced. By using the improved in vitro immunization, induction of B cells secreting antibodies against hydrophilic and hydrophobic regions of TNF-α.
(4)Establishment of CHO cell lines highly producing recombinant human antibodies : By using universal high productive CHO cells, a new system was developed to establish CHO cell lines producing antigen-specific human antibodies in a month.
By developing new technologies for production of human antibodies, the term for developing human antibodies was drastically shortened to several months from several years.
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Report
(4 results)
Research Products
(34 results)
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[Journal Article] IL-10 augments antibody production in in vitro immunized lymphocytes by inducing a Th2-type response and B cell maturation.2004
Author(s)
Xu, Q.H., Katakura, Y, Yamashita, M., Fang, S., Tamura, T., Matsumoto, S., Aiba, Y, Teruya, K, Osada, K, Nishikawa, R., Shirahata, S.
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Journal Title
Bioscience, Biotechnology, and Biochemistry 68
Pages: 2279-2284
NAID
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A hybrid system using both promoter activation and gene amplification for establishing exogenous protein hyper-producing cell lines.2003
Author(s)
Dong, X.Y, Teruya, K, Katakura, Y., Zhang, Y.P., Miura, T., Daimon, Y., Mori, T., Ohashi, H., Shirahata, S.
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Journal Title
Cytotechnology 43
Pages: 11-17
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Construction of an in vitro allergy reaction evaluation system using human leukemia cell lines.2002
Author(s)
Shim, S.Y., Ichikawa, A., Yamashita, M., Katakura, Y., Teruya, K, Mochizuki, Y., Tobinaga, E., Shirahata, S.
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Journal Title
Cytotechnology 40
Pages: 75-83
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Different individual immune responses elicited by in vitro immunization.2002
Author(s)
Yamashita, M., Katakura, Y, Shim, S.Y., Matsumoto, S., Tamura, T., Morihara, K., Aiba, Y, Teruya, K, Tsuchiya, T., Shirahata, S.
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Journal Title
Cytotechnology 40
Pages: 161-165
Description
「研究成果報告書概要(欧文)」より
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