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. 2002 Nov;40(1-3):13-22.
doi: 10.1023/A:1023945517446.

A regulatable selective system facilitates isolation of heterologous protein hyper-producing mammalian cells without gene amplification

Kiichiro Teruya  1 , Yingpei Zhang, Yoshinori Katakura, Sanetaka Shirahata
Affiliations

A regulatable selective system facilitates isolation of heterologous protein hyper-producing mammalian cells without gene amplification

Kiichiro Teruya et al. Cytotechnology. 2002 Nov.

Abstract

In this article, we describe a new method that facilitates to isolate mammalian cells inducible hyper-producing heterologous proteins. This method uses the tetracycline-inducible system to express both the selection marker and the heterologous gene, therefore, allows to increase the selection pressure by reducing the transcription of the selection maker gene. Using this method, we were able to isolate recombinant Chinese hamster ovary cells with a high efficiency. One of established clones produced the recombinant bovine beta-lactoglobulin as heterologous protein at a peak rate of 12 mug 10(-6) cells/day with an inducibility of about 100-fold. This clone was over expressed them RNA of beta-lactoglobulin and the drug resistant gene but did not amplify their genes. When cultured in a hollow fiber bioreactor, the cells were able to secrete beta-lactoglobulinover 300 mug ml(-1). This method is applicable to a broad range of eukaryotic systems and is of general value to technology for recombinant protein production.

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References

    1. Assaraf YG, Schimke RT. Identification of methotrexate transport deficiency in mammalian cells using fluoresceinated methotrexate and flow cytometry. Proc Natl Acad Sci USA. 1987;84:7154–7158. doi: 10.1073/pnas.84.20.7154. - DOI - PMC - PubMed
    1. Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987;7:2745–2752. - PMC - PubMed
    1. Cockett MI, Bebbington CR, Yarranton GT. High level expression of tissue inhibitor of metalloproteinases in Chinese hamster ovary cells using glutamine synthetase gene amplification. Bio/Technology. 1990;8:662–667. doi: 10.1038/nbt0790-662. - DOI - PubMed
    1. Dirks W, Wirth M, Hauser H. Dicistronic transcription units for gene expression in mammalian cells. Gene. 1993;128:247–249. doi: 10.1016/0378-1119(93)90569-O. - DOI - PubMed
    1. Friedman JS, Cofer CL, Anderson CL, Kushner JA, Gray PP, Chapman GE, Stuart MC, Lazarus L, Shine J, Kushner P. High expression in mammalian cells without amplification. Bio/Technology. 1989;8:662–666.