Expression of novel carbohydrate-specific single chain antibodies in E. coli and analyses of carbohydrate-binding activities of purified antibodies
Project/Area Number |
22570125
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Structural biochemistry
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Research Institution | Tokai University |
Principal Investigator |
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Co-Investigator(Renkei-kenkyūsha) |
YAMAGUCHI Yoshiki 独立行政法人理化学研究所, システム糖鎖, チームリーダー (90323451)
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Project Period (FY) |
2010 – 2012
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Project Status |
Completed (Fiscal Year 2012)
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Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2010: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
|
Keywords | 大腸菌発現系 / 単鎖抗体 / 封入体 / リフォールディング / 糖鎖結合性 / 糖鎖親和性 / 昆虫細胞発現系 |
Research Abstract |
Aberration of carbohydrates on cell surface is often associated with pathogenic changes in diseases. Towards developing anti-cancer therapeutics, we had obtained genes for anti-carbohydrate single chain antibodies (scFvs) against carbohydrate moieties expressed specifically in cancer cells using phage-display methods. In this study, expression of scFv genes in E. coli and insect cells was successfully carried out to produce antibody proteins, whose carbohydrate binding activities were confirmed.
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Report
(4 results)
Research Products
(53 results)
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[Journal Article] Effects of two monoclonal antibodies, MLS128 against Tn-antigen and 1H7 against insulin-like growth factor-I receptor, on the growth of colon cancer cells2012
Author(s)
Zamri, N., Masuda, N., Oura, F., Yajima, Y., Nakada, H., and Fujita-Yamaguchi, Y
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Journal Title
Biosci Trends
Volume: 6
Pages: 303-312
Related Report
Peer Reviewed
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[Journal Article] Overproduction of anti-Tn antibody MLS128 single-chain Fv fragment in Escherichia coli cytoplasm using a novel pCold-PDI vector2012
Author(s)
Subedi, G.P., Satoh, T., Hanashima,S., Ikeda, A., Nakada, H., Sato, R., Mizuno M., Yuasa, N., Fujita-Yamaguchi, Y., and Yamaguchi, Y.
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Journal Title
Protein Expr. Puri
Volume: 82
Pages: 197-204
Related Report
Peer Reviewed
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[Journal Article] Construction and expression of anti-Tn-antigen-specific single chain antibody genes from hybridoma producing MLS128 monoclonal antibody2012
Author(s)
Yuasa, N., Ogawa, H., Koizumi, T., Tsukamoto, K., Matsumoto-Takasaki, A., Asanuma, H., Nakada, H., and Fujita- Yamaguchi, Y.
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Journal Title
J Biochem
Volume: 151
Pages: 371-381
Related Report
Peer Reviewed
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[Journal Article] Surface plasmon resonance and NMR analyses of anti Tn-antigen MLS128 monoclonal antibody binding to two or three consecutive Tn-antigen cluster2012
Author(s)
Matsumoto-Takasaki, A., Hanashima, S.. Aoki, A., Yuasa, N., Ogawa, H., Sato, R., Kawakami, H., Mizuno, M. Nakada, H.,Yamaguchi, Y., and Fujita-Yamaguchi, Y
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Journal Title
J Biochem.
Volume: 151
Pages: 273-282
NAID
Related Report
Peer Reviewed
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[Journal Article] Surface plasmon resonance and NMR analyses of anti Tn-antigen MLS128 monoclonal antibody binding to two or three consecutive Tn-antigen cluster2012
Author(s)
Matsumoto-Takasaki, A., Hanashima, S., Aoki, A., Yuasa, N., Ogawa, H., Sato, R., Kawakami, H., Mizuno, M., Nakada, H., Yamaguchi, Y., Fujita-Yamaguchi, Y.
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Journal Title
J Biochem.151
Volume: 151
Pages: 273-282
NAID
Related Report
Peer Reviewed
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[Journal Article] Construction and expression of anti-Tn-antigen-specific single chain antibody genes from hybridoma producing MLS128 monoclonal antibody2012
Author(s)
Yuasa, N., Ogawa, H., Koizumi, T., Tsukamoto, K., Matsumoto-Takasaki, A., Asanuma, H., Nakada, H., Fujita-Yamaguchi, Y.
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Journal Title
J Biochem.151. Feb 8
Volume: 151(Epub ahead of print)(Not known yet)
Related Report
Peer Reviewed
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[Journal Article] Overproduction of anti-Tn antibody MLS128 single-chain Fv fragment in Escherichia coli cytoplasm using a novel pCold-PDI vector2012
Author(s)
Subedi, G.P., Satoh, T., Hanashima, S., Ikeda, A., Nakada, H., Sato, R., Mizuno M., Yuasa, N., Fujita-Yamaguchi Y., Yamaguchi, Y.
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Journal Title
Protein Expr.Puri.
Volume: 82
Pages: 197-204
Related Report
Peer Reviewed
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[Journal Article] Characterization of three different single chain antibodies recognizing nonreducing terminal mannose residues expressed in Escherichia coli by an inducible T7 expression system2011
Author(s)
Matsumoto-Takasaki, A., Yuasa, N., Katagiri, D., Koyama, T., Sakai, K., Zamri, N., Phung, S., Chen, S., Nakada, H., Nakata, M., Fujita-Yamaguchi, Y.
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Journal Title
J Biochem
Volume: 150
Pages: 439-450
Related Report
Peer Reviewed
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[Journal Article] Characterization of three different single chain antibodies recognizing non reducing terminal mannose residues expressed in Escherichia coli by an inducible T7 expression system2011
Author(s)
Matsumoto-Takasaki, A., Yuasa, N., Katagiri, D., Koyama, T., Sakai, K., Zamri, N., Phung, S., Chen, S., Nakada, H., Nakata, M., Fujita-Yamaguchi, Y.
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Journal Title
J Biochem.150
Volume: 150
Pages: 439-450
Related Report
Peer Reviewed
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[Journal Article] Production of anti-carbohydrate antibodies by phage-display technologies: Potential impairment of cell growth as a result of endogenous expression2010
Author(s)
Yuasa, N., Zhang, W., Goto, T., Sakaue, H., Matsumoto- Takasaki, A., Kimura, M., Ohshima, H., Tsuchida, Y., Koizumi, T., Sakai, K., Kojima, T., Yamamoto, K., Nakata, M., and Fujita-Yamaguchi, Y.
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Journal Title
J. Biol. Chem
Volume: 285
Pages: 30587-30597
Related Report
Peer Reviewed
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[Journal Article] Production of anti-carbohydrate antibodies by phage-display technologies : Potential impairment of cell growth as a result of endogenous expression.2010
Author(s)
Yuasa, N., Zhang, W., Goto, T., Sakaue, H., Matsumoto- Takasaki, A., Kimura, M., Ohshima, H., Tsuchida, Y., Koizumi, T., Sakai, K., Kojima, T., Yamamoto, K., Nakata, M., Fujita-Yamaguchi, Y.
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Journal Title
J.Biol.Chem.
Volume: 285
Pages: 30587-30597
Related Report
Peer Reviewed
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[Presentation] NMR analysis of an anti-carbohydrate antibody MLS128 single-chain Fv fragment toward elucidation of the multivalent recognition mechanism2011
Author(s)
Subedia, G.P., Satoh, T., Hanashima, S., Ikeda, A., Nakada, H., Sato, R., Mizuno, M., Yuasae, N., Fujita-Yamaguchi, Y., Yamaguchi, Y.
Organizer
The 50th Memorial Annual Meeting of the Nuclear Magnetic Resonance Society of Japan
Place of Presentation
Yokohama (Osanbashi Hall)
Related Report
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[Presentation] Establishment of overproduction procedure for anti-Tn antigen MLS128 single-chain Fv fragment toward the structural studies2010
Author(s)
Subedi, G. P., Satoh, T., Hanashima, S., Ikeda, A, Nakada, H., Sato, R., Mizuno, M., Yuasa, N., Fujita-Yamaguchi, Y., and Yamaguchi, Y.
Organizer
BMB2010 (日本生化学会・分子生物学会)
Place of Presentation
神戸ポートアイランド(兵庫県)
Related Report
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[Presentation] In vitro production of anti-carbohydrate antibodies : Purification of anti-T-antigen (beta Gall-3GalNAc-Thr/Ser) single chain antibodies (scFvs) from inclusion bodies.2010
Author(s)
Matsumoto-Takasaki, A., Yamaguchi, S., Koizumi, T., Yuasa, N., Fujita-Yamaguchi, Y.
Organizer
240^<th> American Chemical Society National Meeting and Exposition
Place of Presentation
Boston, MA, USA
Related Report
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[Presentation] Refolding and characterization of three anti-mannotriose single chain antibodies2010
Author(s)
Katagiri, D., Matsumoto-Takasaki, A., Yuasa, N., Koyama, T., Ogawa, H. Sakai, K., Takayanagi, A., Nakata, M., Fujita-Yamaguchi, Y.
Organizer
BMB2010(日本生化学会・分子生物学会)
Place of Presentation
神戸ポートアイランド(兵庫県)
Related Report
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[Presentation] Purification and characterization of anti-LNFPIII antibody expressed in CHO cells2010
Author(s)
Tsutida, Y., Yuasa, N., Goto, T., Kubo, T., Zhang, W., Fujita-Yamaguchi, Y.
Organizer
BMB2010(日本生化学会・分子生物学会)
Place of Presentation
神戸ポートアイランド(兵庫県)
Related Report
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[Presentation] Establishment of overproduction procedure for anti-Tn antigen MLS128 single-chain Fv fragment toward the structural studies.2010
Author(s)
Subedi, G. P., Satoh, T., Hanashima, S., Ikeda, A, Nakada, H., Sato, R., Mizuno, M., Yuasa, N., Fujita-Yamaguchi, Y., Yamaguchi, Y.
Organizer
BMB2010(日本生化学会・分子生物学会)
Place of Presentation
神戸ポートアイランド(兵庫県)
Related Report