N-terminal vacuolar sorting signal at the mouse antibody alters the N-linked glycosylation pattern in suspension-cultured tobacco BY2 cells(PLANT BIOTECHNOLOGY)
スポンサーリンク
概要
- 論文の詳細を見る
Recombinant DNA technology enables the use of plants as the host for the production of pharmaceutical proteins, such as antibodies. The glycosylation of recombinant proteins plays physiological and biological roles. However, because glycosylation in plants is different from that in human cells, the development of glycoengineering is required. In plant cells, glycan structures are shown to correlate with the localization of the recombinant protein produced. In this study, the vacuolar sorting signal (VSS) of sporamin was fused to the heavy (H) and light (L) chains of a mouse monoclonal antibody (mAb), and the mAb was produced in suspension-cultured tobacco BY2 cells. The sugar chain structures were determined by high-performance liquid chromatography, exoglycosidase digestion, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Typical plant glycans with α1,3-fucosylation and/or β1,2-xylosylation derived from mAb with the VSS-fused H-chain (mIgG1000) and mAb with the VSS-fused H- and L-chain (mIgG1010) occupied the large amount of the total N-glycans, 72.1% and 85.0%, respectively, such as those derived from mAb without VSS (mIgG0000), 74.6% (Fujiyama et al., J. Biosci. Bioeng., 101, 212-218, 2006). In contrast, the typical plant glycan structure Man_3FucXylGlcNAc_2 particularly in vacuoles accounted for 37.8% of the total sugar chains derived from mIgG1000 and 58.5% of those derived from mIgG1010 compared with 24.3% of those derived from mIgG0000. These results suggest that the sporamin signal peptide fused to mAb acts as a VSS and leads to the increase in the amount of Man_3FucXylGlcNAc_2, which is the main N-glycan structure in vacuoles.
著者
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Fujiyama Kazuhito
International Center for Biotechnology, Osaka University
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大政 健史
大阪大学大学院工学研究科生命先端工学専攻
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大政 健史
大阪大学大学院工学研究科
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Omasa Takeshi
大阪大学生物工学国際交流センター
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Omasa Takeshi
大阪大学 工研究
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Omasa Takeshi
Grad. Sch. Of Eng. Osaka Univ.
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Ohya Tomoshi
Mitsubishi Tanabe Pharma Corporation
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SEKI Tatsuji
International Center for Biotechnology, Osaka University
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Kazuhito Fujiyama
International Center For Biotechnology Osaka University
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Fujiyama Kazuhito
大阪大学生物工学国際交流センター
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Fujiyama Kazuhito
International Center For Biotechnology Osaka Univ.
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Fujiyama Kazuhito
Department Of Fermentation Technology Osaka University
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Fujiyama Kazuhito
International Center For Biotechnology Osaka University
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Misaki Ryo
International Center For Biotechnology Osaka University
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Misaki Ryo
The International Center For Biotechnology Osaka University
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Seki Tatsuji
International Center For Biotechnology Osaka University
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Seki Tatsuji
大阪大学生物工学国際交流センター
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Takami Takashi
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Omasa Takeshi
大阪大学 工学研究科生物工学
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Sakai Yohei
International Center For Biotechnology Osaka University
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Fujiyama Kazuhito
The International Center For Biotechnology Osaka Univ. 2-1 Yamada-oka Suita Osaka 565-0871 Jpn
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大政 健史
徳大院
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Omasa Takeshi
Department Of Biological Science And Technology Institute Of Technology And Science The University O
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Omasa Takeshi
Department Of Biological Science And Technology Biosystems Engineering Institute Of Technology And Science The University Of Tokushima
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大政 健史
徳大院・STS研:阪大院・工・生命先端
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大政 健史
徳大院・STS研:阪大院・工
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