Efficient production of single-chain Fv antibody possessing rare codon linkers in fed-batch fermentation(BIOCHEMICAL ENGINEERING)
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概要
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Single-chain Fv antibody (scFv) having 2 types of polypeptide linkers with or without rare codons, namely scFv (G_4S)^R_3 and scFv No. 10 (with rare codons) and scFv (G_4S)_3 and scFv No. 10^<NR> (without rare codon), were expressed under controllable conditions in batch and fed-batch fermentation, in order to compare volumetric productivity and specific productivity levels of scFvs as a soluble form. In batch fermentation, volumetric productivity levels of scFv (G_4S)^R_3 and scFv No. 10, namely scFvs having the rare codon linkers were 3-5 times higher than those of scFvs that had linkers without the rare codon. In fed-batch fermentation controlled by an exponential feeding system, the cell concentrations of the transformants increased with similar specific growth rates (0.1h^<-1>), while the specific productivity levels of scFvs with the rare codon linkers were 1.6 times higher than those of scFvs without the rare codon linkers. These results indicate that the presence of several rare codons in the gene of a polypeptide linker increases soluble amount of scFvs. This might be caused by a temporary decrease in translation speed at the position of the polypeptide linker allowing time for the folding of the V_H domain and avoiding unfavorable interactions between amino acid residues at the unfolded V_H and V_L domains. Higher specific productivity levels of both scFv No. 10 and scFv No. 10^<NR> than those of scFv (G_4S)^R_3 and (G_4S)_3 might be caused by difference in stability of the polypeptide linkers on the basis of amino acid sequences. Thus, the rare codon linkers tested in this study will be considerably useful for large-scale production of soluble and active scFvs in fed-batch or continuous fermentations, in which high cell activity can be maintained.
- 公益社団法人日本生物工学会の論文
著者
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Yamaji Hideki
Graduate School Of Engineering Department Of Chemical Science And Engineering Kobe University
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Kumada Yoichi
Department Of Bioscience And Biotechnology Okayama University
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Seong Gi
Department Of Applied Chemistry Hanyang University
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Katoh Shigeo
Graduate School Of Engineering Department Of Chemical Science And Engineering Kobe University:depart
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Kikuchi Yasufumi
Graduate School Of Engineering Department Of Chemical Science And Engineering Kobe University
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Sakan Yoshinobu
Graduate School of Engineering, Department of Chemical Science and Engineering, Kobe University
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Kajihara Hideyuki
Graduate School of Engineering, Department of Chemical Science and Engineering, Kobe University
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Kihara Mana
Graduate School of Engineering, Department of Chemical Science and Engineering, Kobe University
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Kihara Mana
Graduate School Of Engineering Department Of Chemical Science And Engineering Kobe University
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Sakan Yoshinobu
Graduate School Of Engineering Department Of Chemical Science And Engineering Kobe University
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Kajihara Hideyuki
Graduate School Of Engineering Department Of Chemical Science And Engineering Kobe University
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