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Long-term operation of depth filter perfusion systems (DFPS) for monoclonal antibody production using recombinant CHO cells: Effect of temperature, pH, and dissolved oxygen


Recombinant CHO cells of DG44 origin (CS*13-1.00), expressing a chimeric antibody against the S surface antigen of the Hepatitis B virus, were cultivated in single-stage and two-stage depth filter perfusion systems (DFPS) under varying temperature, pH, and oxygen tension conditions to determine their effects on recombinant antibody production. A long-term culture was carried out in a single-stage depth filter for 81 days, during which an occasional clog interrupted the experiment. However, this problem was solved via trypsin injection. The DFPS showed a steady production of monoclonal antibody at a concentration of 100∼150 mg/L. As the cultivation temperature was increased from 33 to 37°C, the monoclonal antibody (Mab) concentration increased from 80.33 to 133.47 mg/L. Likewise, the glucose uptake rate (GUR) and lactate production rate (LPR) also increased. With an increase in pH from 6.95 to 7.61, the Mab concentration increased from 61.64 to 94.31 mg/L. When the oxygen tension was increased from 60 to 80%, the Mab concentration increased from 93.78 to 128.30 mg/L.

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  1. Voisard, D., F. Meuwly, P.-A. Ruffieux, G. Baer, and A. Kadouri (2003) Potential of cell retention techniques for large-scale high-density perfusion culture of suspended mammalian cells. Biotechnol. Bioeng. 82: 751–765.

    Article  CAS  Google Scholar 

  2. Oh, D. J., S. K. Choi, and H. N. Chang (1994) Highdensity continuous culture of hybridoma cells in a depth filter perfusion system. Biotechnol. Bioeng. 44: 895–901.

    Article  CAS  Google Scholar 

  3. Choi, S. K., H. N. Chang, G. M. Lee, I. H. Kim, and D. J. Oh (1995) High cell density perfusion cultures of anchorage-dependent Vero cells in a depth filter perfusion system. Cytotechnology 17: 173–183.

    Article  CAS  Google Scholar 

  4. Choi, S. K., H. N. Chang, and D. J. Oh (1995) Continuous production of tissue plasminogen activator from recombinant CHO cells in a depth filter perfusion system. Biotechnol. Tech. 9: 567–572.

    Article  Google Scholar 

  5. Lee, J. C., H. N. Chang, and D. J. Oh (2005) Recombinant antibody production by perfusion cultures of rCHO cells in a depth filter perfusion system. Biotechnol. Prog. 21: 134–139.

    Article  CAS  Google Scholar 

  6. Kwon, S., I. K. Yoo, W. G. Lee, H. N. Chang, and Y. K. Chang (2001) High-rate continuous production of lactic acid by Lactobacillus rhamnosus in a two-stage membrane cell-recycle bioreactor. Biotechnol. Bioeng. 73: 25–34.

    Article  CAS  Google Scholar 

  7. Lee, J. C., D. Y. Kim, D. J. Oh, and H. N. Chang (2008) Two-stage depth filter perfusion culture for recombinant antibody production by recombinant Chinese hamster ovary cell. Biotechnol. Bioprocess Eng. In press.

  8. Yoon, S. K., J. Y. Song, and G. M. Lee (2003) Effect of low culture temperature on specific productivity, transcription level, and heterogeneity of erythropoietin in Chinese hamster ovary cells. Biotechnol. Bioeng. 82: 289–298.

    Article  CAS  Google Scholar 

  9. Yoon, S. K., S. L. Choi, J. Y. Song, and G. M. Lee (2005) Effect of culture pH on erythropoietin production by Chinese hamster ovary cells grown in suspension at 32.5 and 37°C. Biotechnol. Bioeng. 89: 345–356.

    Article  CAS  Google Scholar 

  10. Simpson, N. E., L. K. Bennett, K. K. Papas, A. Sambanis, and I. Constantinidis (2000) Effects of pH on murine insulinoma βTC3 cells. Biochem. Biophys. Res. Commun. 273: 937–941.

    Article  CAS  Google Scholar 

  11. Zhang, F., M. A. Saarinen, L. J. Itle, S. C. Lang, D. W. Murhammer, and R. J. Linhardt (2002) The effect of dissolved oxygen (DO) concentration on the glycosylation of recombinant protein produced by the insect cell-baculovirus expression system. Biotechnol. Bioeng. 77: 219–224.

    Article  CAS  Google Scholar 

  12. Taticek, R. A. and M. L. Shuler (1997) Effect of elevated oxygen and glutamine levels on foreign protein production at high cell densities using the insect cellbaculovirus expression system. Biotechnol. Bioeng. 54: 142–152.

    Article  CAS  Google Scholar 

  13. Ryll, T., G. Dutina, A. Reyes, J. Gunson, L. Krummen, and T. Etcheverry (2000) Performance of small-scale CHO perfusion cultures using an acoustic cell filtration device for cell retention: Characterization of separation efficiency and impact of perfusion on product quality. Biotechnol. Bioeng. 69: 440–449.

    Article  CAS  Google Scholar 

  14. Jenkins, N. and A. Hovey (1993) Temperature control of growth and productivity in mutant Chinese hamster ovary cells synthesizing a recombinant protein. Biotechnol. Bioeng. 42: 1029–1036.

    Article  CAS  Google Scholar 

  15. Kim, S. J., N. S. Kim, C. J. Ryu, H. J. Hong, and G. M. Lee (1998) Characterization of chimeric antibody producing CHO cells in the course of dihydrofolate reductase-mediated gene amplification and their stability in the absence of selective pressure. Biotechnol. Bioeng. 58: 73–84.

    Article  CAS  Google Scholar 

  16. Chang, H. N. and M. Moo-Young (1988) Analysis of oxygen transport in immobilized whole cells. pp. 33–51. In: M. Moo-Young (ed.). Bioreactor Immobilized Enzymes and Cells: Fundamentals and Applications. Elsevier Pub., The Netherlands.

    Google Scholar 

  17. Kim, D. Y. (2006) Process Development for Production of Recombinant Antibody by rCHO Cells. Ph.D. Thesis. KAIST, Daejeon, Korea.

    Google Scholar 

  18. Chang, H. N., B. J. Kim, J. W. Kang, C. M. Jeong, N. J. Kim, and J. K. Park (2008) High cell density ethanol fermentation in an upflow packed-bed cell recycle bioreactor. Biotechnol. Bioprocess Eng. 13: 123–135.

    Article  CAS  Google Scholar 

  19. Kim, B. J., H. N. Chang, and D. J. Oh (2007) Application of a cell-once-through perfusion strategy for production of recombinant antibody from rCHO cells in a Centritech Lab II centrifuge system. Biotechnol Prog. 23: 1186–1197.

    CAS  Google Scholar 

  20. Kim, B. J., D. J. Oh, and H. N. Chang (2008) Limited use of Centritech Lab II Centrifuge in perfusion culture of rCHO cells for the production of recombinant antibody. Biotechnol. Prog. 24: 166–174.

    Article  CAS  Google Scholar 

  21. Meuwly, F., U. Weber, T. Ziegler, A. Gervais, R. Mastrangeli, C. Crisci, M. Rossi, A. Bernard, U. von Stockar, and A. Kadouri (2006) Conversion of a CHO cell culture process from perfusion to fed-batch technology without altering product quality. J. Biotechnol. 123: 106–116.

    Article  CAS  Google Scholar 

  22. Wang, M. D., M. Yang, N. Huzel, and M. Butler (2002) Erythropoietin production from CHO cells grown by continuous culture in a fluidized-bed bioreactor. Biotechnol. Bioeng. 77: 194–203.

    Article  CAS  Google Scholar 

  23. Zhou, W., J. Rehm, A. Europa, and W. S. Hu (1997) Alternation of mammalian cell metabolism by dynamic nutrient feeding. Cytotechnology 24: 99–108.

    Article  CAS  Google Scholar 

  24. Warnock, J. N. and M. Al-Rubeai (2006) Bioreactor systems for the production of biopharmaceuticals from animal cells. Biotechnol. Appl. Biochem. 45: 1–12.

    Article  CAS  Google Scholar 

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Correspondence to Duk Jae Oh or Ho Nam Chang.

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Lee, J.C., Kim, D.Y., Oh, D.J. et al. Long-term operation of depth filter perfusion systems (DFPS) for monoclonal antibody production using recombinant CHO cells: Effect of temperature, pH, and dissolved oxygen. Biotechnol Bioproc E 13, 401–409 (2008).

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  • Chinese hamster ovary cells
  • perfusion culture
  • recombinant antibody
  • two-stage depth filter perfusion system