High-level expression of recombinant IgG1 by CHO K1 platform

Ningning Xu, Jianfa Ou, Al-Karim (Al) Gilani, Lufang Zhou, Margaret Liu

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PDF(382 KB)
Front. Chem. Sci. Eng. ›› 2015, Vol. 9 ›› Issue (3) : 376-380. DOI: 10.1007/s11705-015-1531-5
RESEARCH ARTICLE
RESEARCH ARTICLE

High-level expression of recombinant IgG1 by CHO K1 platform

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Abstract

The Chinese Hamster Ovary (CHO K1) cell was used to express a targeted anti-cancer monoclonal antibody by optimizing the platform of the construction of production cell line in this study. The adherent CHO K1 was first adapted to suspension culture in chemical defined medium. Then the glutamine synthetase (GS) vector was applied to construct a single plasmid to overexpress a monoclonal antibody IgG1. Post transfection, the production of cell pool was optimized by glutamine-free selection and amplification using various concentrations of methionine sulfoximine. The best cell pool of CHO K1/IgG1 was used to screen the top single clone using the limiting dilution cloning. Finally, a high IgG1 production of 780 mg/L was obtained from a batch culture. This study demonstrated that the construction of high producing cell line, from gene to clone, could be completed within six month and the gene amplification improved protein production greatly.

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Keywords

Chinese hamster ovary (CHO) / monoclonal antibody / IgG1 / amplification / cell line development

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Ningning Xu, Jianfa Ou, Al-Karim (Al) Gilani, Lufang Zhou, Margaret Liu. High-level expression of recombinant IgG1 by CHO K1 platform. Front. Chem. Sci. Eng., 2015, 9(3): 376‒380 https://doi.org/10.1007/s11705-015-1531-5

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Acknowledgements

This work was supported by RGC fund from The University of Alabama and the BRIGE grant (No. 1342390 and IIP-1026648) from the National Science Foundation.

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg
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