Long-term operation of depth filter perfusion systems (DFPS) for monoclonal antibody production using recombinant CHO cells: Effect of temperature, pH, and dissolved oxygen Authors
First Online: 20 September 2008 Received: 30 May 2008 Accepted: 24 June 2008 DOI:
Cite this article as: Lee, J.C., Kim, D.Y., Oh, D.J. et al. Biotechnol Bioproc E (2008) 13: 401. doi:10.1007/s12257-008-0155-8 Abstract
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.
Keywords Chinese hamster ovary cells perfusion culture recombinant antibody two-stage depth filter perfusion system Download to read the full article text References
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.
Oh, D. J., S. K. Choi, and H. N. Chang (1994) Highdensity continuous culture of hybridoma cells in a depth filter perfusion system.
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.
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.
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.
Kwon, S., I. K. Yoo, W. G. Lee, H. N. Chang, and Y. K. Chang (2001) High-rate continuous production of lactic acid by
in a two-stage membrane cell-recycle bioreactor.
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.
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.
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.
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.
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.
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.
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.
Jenkins, N. and A. Hovey (1993) Temperature control of growth and productivity in mutant Chinese hamster ovary cells synthesizing a recombinant protein.
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.
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.
Kim, D. Y. (2006)
Process Development for Production of Recombinant Antibody by rCHO Cells. Ph.D. Thesis. KAIST, Daejeon, Korea.
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.
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.
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.
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.
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.
Zhou, W., J. Rehm, A. Europa, and W. S. Hu (1997) Alternation of mammalian cell metabolism by dynamic nutrient feeding.
Warnock, J. N. and M. Al-Rubeai (2006) Bioreactor systems for the production of biopharmaceuticals from animal cells.
Biotechnol. Appl. Biochem.
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