Abstract
This report shows a method to establish a recombinant CHO cell line that grows and secretes human antithrombin-III (AT-III) in a serum-free medium. At first, serum-dependent cell lines were obtained by introducing AT-III cDNA and following MTX-treatments. Then the cells were transferred to a serum-free medium, GCM001, and cultivated for several weeks. Serum- independent cells were obtained through cultivation in GCM001, and some cell lines showed a little greater productivity of AT-III in GCM001 than serum-dependent, parental cells in serum-containing medium. AT-III produced by both serum-independent and serum-dependent cells had the same specific activity, but heterogeneity in glycosylations was observed between them.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Wasley L.C., Atha, D.H., Bauer, K.A. and Kaufman R.J. (1987) ‘Expression and Characterization of Human Antithrombin-III Synthesized in mammalian cells’, J. Biol. Chem, 262, 14766–14772.
Zettlmeissl, G., Ragg, H. and Karges, H.E. (1987) ‘Expression of Biologically Active Human Antithrombiniii in Chinese Hamster Ovary Cells,’ Biotechnology, 5, 720–725
Bock S.C., Wion, K.L., Vehar, G.A. and Lawn, R.M. (1982) ‘Cloning andexpression of the cDNA for human antithrombin III’, Nucleic Acids Res., 10, 8113–8125
Lee, F., Mulligan, R., Berg, P. and Ringold, G. ‘Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumor virus chimeric plasmids’, Nature, 294, 228–232
Ricco, A., Grimaldi, G., Verde, P., Sebastio, G., Boast, S. and Blasi, F. (1985) ‘The human urokinase-plasminogen activator gene and promoter,’ Nucleic Acids Res., 13, 2759–2771
Urlaub, G. and Chaisin, L.A. (1980) ‘Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity’, Proc. Natl. Acad. Sci. USA/77, 4216–4220
Gorman, C. (1985) ‘High efficiency gene transfer into mammalian cells’, in Glover, D.M. (eds.), DNA cloning vol. II, IRL Press, Washington, pp.143–190
Molly, C. Owen, M.C. and Boswell D.R. (1989) ‘Immunoadsorbent Purification of Antithrombin;Active form and Inactive Variants’, Thromb. Res., 55, 657–660
Laemmli, U.K. (1970) ‘Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4’, Nature, 227, 680–685
Yamane, I. and Murakami, K. (1971) ‘A New Crystalline Growth Factor for Mammalian Cells in Vitro, Isolated from a Commecial Peptone’, Biochem. Biophys. Res. Commun., 45, 1431–1436
Keay, L. (1976) ‘Autoclavable Low Cost Serum-Free Cell Culture Media; The Growth of Established Cell Lines and Production of Viruses’, Biotech. Bioeng., 18, 363–382
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1992 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Asakura, E. et al. (1992). Production of Human Antithrombin-III in a Serum-Free Culture of CHO Cells. In: Murakami, H., Shirahata, S., Tachibana, H. (eds) Animal Cell Technology: Basic & Applied Aspects. Animal Cell Technology: Basic & Applied Aspects, vol 4. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-2844-5_47
Download citation
DOI: https://doi.org/10.1007/978-94-011-2844-5_47
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-5263-4
Online ISBN: 978-94-011-2844-5
eBook Packages: Springer Book Archive