High-level expression of recombinant IgG1 by CHO K1 platform

doi:10.1007/s11705-015-1531-5

Front. Chem. Sci. Eng.

2015, Vol. 9

Issue (3) : 376-380 https://doi.org/10.1007/s11705-015-1531-5

RESEARCH ARTICLE

High-level expression of recombinant IgG1 by CHO K1 platform

Ningning Xu¹,Jianfa Ou¹,Al-Karim (Al) Gilani¹,Lufang Zhou^2,^*(

),Margaret Liu¹

1. Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL 35294, USA
2. Departments of Medicine and Biomedical Engineering, The University of Alabama at Birmingham, Birmingham, AL 35294, USA

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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.

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

Corresponding Author(s): Lufang Zhou

Online First Date: 24 September 2015 Issue Date: 30 September 2015

Cite this article:

Ningning Xu,Jianfa Ou,Al-Karim (Al) Gilani, et al. High-level expression of recombinant IgG1 by CHO K1 platform[J]. Front. Chem. Sci. Eng., 2015, 9(3): 376-380.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-015-1531-5
https://academic.hep.com.cn/fcse/EN/Y2015/V9/I3/376

Fig.1 pEE-IgG plasmid construction (left). hCMV-MIE: human cytomeg-alovirus major immediate-early enhancer and promoter; SV40 ori: SV40 origin of replication; GS: glutamine synthetase. Plasmid analysis by DNA gel electrophoresis (right). Lane 1: DNA marker; Lane 2: pEE-IgG

Fig.2 Cell growth kinetics of CHO K1/IgG cell pool

Fig.3 IgG Production of top 6 clones in shaker flask batch culture

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