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Efficient production of single-chain Fv antibody possessing rare codon linkers in fed-batch fermentation

Paper ID Volume ID Publish Year Pages File Format Full-Text
21959 43245 2009 5 PDF Available
Title
Efficient production of single-chain Fv antibody possessing rare codon linkers in fed-batch fermentation
Abstract

Single-chain Fv antibody (scFv) having 2 types of polypeptide linkers with or without rare codons, namely scFv (G4S)3R and scFv No.10 (with rare codons) and scFv (G4S)3 and scFv No.10NR (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 (G4S)3R 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 (0.1 h− 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 VH domain and avoiding unfavorable interactions between amino acid residues at the unfolded VH and VL domains. Higher specific productivity levels of both scFv No. 10 and scFv No. 10NR than those of scFv (G4S)3R and (G4S)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.

Keywords
scFv; Rare codon linker; Specific production rate; Fed-batch fermentation; Exponential feeding system
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Efficient production of single-chain Fv antibody possessing rare codon linkers in fed-batch fermentation
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 107, Issue 1, January 2009, Pages 73–77
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us