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Cytotechnology

, Volume 54, Issue 2, pp 89–96 | Cite as

Enhanced recombinant M-CSF production in CHO cells by glycerol addition: model and validation

  • Chi-Hsien LiuEmail author
  • Li-Hsin Chen
Original Research

Abstract

Addition of stimulatory chemical such as glycerol was found to increase recombinant protein production in Chinese hamster ovary (CHO) cells. However, glycerol influenced cell mitosis and reduced cell growth rate. We developed a controlled proliferation strategy to utilize the stimulation of glycerol on recombinant protein production and mitigate the problem of growth inhibition. The approach is to apply a two-stage process, where cells are cultured without glycerol for a period of time in order to obtain enough cell density and then glycerol is added to achieve high specific productivity. In addition, a model for predicting the profiles of cell proliferation and recombinant protein production was developed and validated. A two-stage process, addition of 1% glycerol after 1 day of growth, could increase the final production of macrophage-colony stimulating factor (M-CSF) by 38% compared with the value obtained without addition of glycerol.

Keywords

CHO cells Controlled proliferation Glycerol M-CSF Model Optimization 

Notes

Acknowledgements

The authors were supported by National Science Council (94-2214-E-182-008) and Chang Gung Memorial Hospital (CMRPD32051).

References

  1. Arden N, Betenbaugh AMJ (2006) Regulating apoptosis in mammalian cell cultures. Cytotechnology 50:77–92CrossRefGoogle Scholar
  2. Brown CR, Hong-Brown LQ, Biwersi J, Verkman AS, Welch WJ (1996) Chemical chaperones correct the mutant phenotype of the delta F508 cystic fibrosis transmembrane conductance regulator protein. Cell Stress Chaperones 1:117–125CrossRefGoogle Scholar
  3. Fagain CO (1997) Use of stabilizing additives. in stabilizing protein function. Springer, New York, pp. 70–79Google Scholar
  4. Fox SR, Patel UA, Yap MGS, Wang DIC (2004) Maximizing interferon-γ production by Chinese hamster ovary cells through temperature shift optimization: experimental and modeling. Biotechnol Bioeng 85:177–184CrossRefGoogle Scholar
  5. Fussenegger M, Bailey JE (1999) Control of mammalian cell proliferation as an important strategy in cell culture technology, cancer therapy and tissue engineering. In: Al–Rubeai M, Hauser H, Jenkins N, Betenbaugh MJ, McDonald (eds) Cell engineering I. Kluwer Academic, Norwell, MA, pp. 186– 219Google Scholar
  6. Gorovits BM, McGee WA, Horowitz PM (1998) Rhodanese folding is controlled by the partitioning of its folding intermediates. Biochim Biophys Acta 1382:120–128Google Scholar
  7. Jaspard E (2000) Role of protein-solvent interactions in refolding: effects of cosolvent additives on the renaturation of porcine pancreatic elastase at various pHs. Arch Biochem Biophys 375:220–228CrossRefGoogle Scholar
  8. Kim DY, Lee JC, Chang HN, Oh DJ (2005) Effects of supplementation of various medium components on Chinese hamster. Cytotechnology 47:37–49CrossRefGoogle Scholar
  9. Li G., Hong YH, Wu KF, Lin YM, Cao ZY, Zheng GG (2002) Clone and expression of mutant M-CSF and its receptor from human leukemic cell line J6–1. Leukemia Res 26:377–382CrossRefGoogle Scholar
  10. Mishra R, Seckler R, Bhat R (2005) Efficient refolding of aggregation-prone citrate synthase by polyol osmolytes: how well are protein folding and stability aspects coupled. J Biol Chem 280:15553–15560CrossRefGoogle Scholar
  11. Nienow AW (2006) Reactor engineering in large scale animal cell culture. Cytotechnology 50:9–33CrossRefGoogle Scholar
  12. Rodriguez J, Spearman M, Huzel N, Butler M (2005) Enhanced production of monomeric interferon by CHO cells through the control of culture conditions. Biotechnol Prog 21:22–30CrossRefGoogle Scholar
  13. Shuler ML, Kargi F (2002) Bioprocess engineering- basic concepts. Prentice Hall Inc. Upper Saddle River, NJ, USAGoogle Scholar
  14. Tieman BC, Johnston MF, Fisher MT (2001) A comparison of the GroE chaperonin requirements for sequentially and structurally homologous malate dehydrogenases. J Biol Chem 276:44541–44550CrossRefGoogle Scholar
  15. Timasheff SN (2002) Protein hydration, thermodynamic binding, and preferential hydration. Biochemistry 41:13473–13482CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Graduate Institute of Biochemical and Biomedical EngineeringChang Gung UniversityTaoyuanTaiwan

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