Functional Construction of the Anti-Mucin Core Protein (MUC1) Antibody MUSE11 Variable Regions in a Bacterial Expression System.
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概要
- 論文の詳細を見る
A bacterial expression system for the variable region fragments (Fvs) of the anti-MUC 1 tumor antigen antibody MUSE 11 has been constructed. The Fv fragment showed binding specificity toward TFK-1 cells, with slightly reduced affinity compared to its parent IgG. The single-chain Fv fragment was arranged in two orders, VH-linker-VL and VL-linker-VH. However, linking the regions with a flexible peptide linker (GGGGS), or with a shorter linker (GGGGS) led to a dramatic decrease in the biological activity toward the target antigen in both arrangements, suggesting that the MUSE 11 antibody loses its activity when the domains are linked with polypeptide linkers. These results indicate that the variable region domains of the anti-MUC 1 antibody MUSE 11 have specificity only in the Fv form, and that linking the domains strongly reduces the association with its target antigen. Gel filtration analysis indicates that the scFv has a dimeric structure, suggesting that the inactivation of MUSE 11 scFv is due to unfavorable intermolecular associations of the scFv chains. To our knowledge, this is the first report of a significant reduction in affinity caused by linking the variable domains in both arrangements, i.e., VH-VL and VL-VH.
- 社団法人 日本生化学会の論文
著者
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Matsuno Seiki
First Department Of Surgery Tohoku University School Of Medicine
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Imai Kohzoh
Deparment Of Internal Medicine Sapporo Medical College
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Tsumoto Kouhei
Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University
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SUZUKI Masanori
First Department of Surgery, Tohoku University School of Medicine
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Kumagai Izumi
Department Of Biochemistry And Engineering Graduate School Of Engineering Tohoku University
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Kudo Toshio
Cell Resource Center For Biomedical Research Institute Of Development Aging And Cancer Tohoku Univer
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Asano Ryutaro
Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University
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Shinoda Masao
First Department Of Surgery Tohoku University School Of Medicine
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KATAYOSE Yu
Cell Resource Center for Biomedical Research, Institute of Development, Aging and Cancer, Tohoku Uni
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Ebara Shinji
Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University
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Takemura Shin-ichi
Department Of Applied Chemistry Faculty Of Engineering Kyushu Institute Of Technology
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Sakurai Naoki
Department Of Biology
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Teramae Atsushi
Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University
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