Production of bioactive trastuzumab and chimeric anti-VEGF antibody in the cytoplasm of Escherichia coli
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Author list: Charoenpun P., Leelawattanachai J., Meechai A., Waraho-Zhmayev D.
Publisher: IOP Publishing
Publication year: 2018
Volume number: 185
Issue number: 1
ISSN: 1755-1307
eISSN: 1755-1315
Languages: English-Great Britain (EN-GB)
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Abstract
Monoclonal antibody (mAb) is a very useful tool in not only medical field but also in research field. However, the current cost of its production is very high as most mAbs are produced in mammalian cell system. Escherichia coli is a good alternative but has many bottlenecks. Recently, it was demonstrated that biological active full-length Immunoglobulin G (IgG) could be produced in the reductive cytoplasm of an engineered E. coli strain called SHuffle. In this study, we investigated the effect of temperature and induction level on the production of full-length humanized anti-Her2 and chimeric anti-VEGF IgGs expressed as cyclonal. The solubility as well as the assembly of the IgG molecules was examined by western blot analysis. It was found that expression at 30 ฐC with 0.1 mM IPTG induction was the most suitable for trastuzumab. In contrast, the full-length expression of chimeric anti-VEGF required induction with 1 mM IPTG at 30 ฐC., however, 1 mM IPTG induction resulted in more full-length IgG. In conclusion, the present study indicated that E coli could be successfully used for the full-length expression of IgG by suitably modifying the expression conditions. ฉ 2018 Institute of Physics Publishing. All rights reserved.
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