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The influence of dissolved oxygen tension on the metabolic activity of an immobilized hybridoma population

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Abstract

In the study presented here a laboratory scale (150 ml) fluidized bed bioreactor was used as a tool for making kinetic measurements on the production of monoclonal antibodies (MAb) with a hybridoma cell line. We determined the influence of dissolved oxygen tension (DOT) on the metabolic activity of a hybridoma population immobilized in macroporous collagen microspheres. The data obtained showed a reduction of the metabolic activity of the immobilized population at reduced DOT, the total number of immobilized cells, however, remained constant. At decreasing DOT an increasing lactate yield from glucose at reduced glutamine consumption was noticed, indicating a shift in the pattern of substrate usage. A mathematical description of maintenance metabolism was formulated and the parameters of growth and maintenance requirements were calculated. A growth associated MAb production was determined under the conditions applied leading to space time yields of 225 mg MAb per liter of total reactor volume and day.

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References

  • Almgren J, Nilsson C, Petersson AC and Nilsson, K (1991) Cultispher—macroporous gelatin microcarrier—new applications. In: Spier RE, Griffiths JB and Meignier B (eds) Production of Biologicals from Animal Cells in Culture (pp. 434–438) Oxford, Butterworth-Heinemann.

    Google Scholar 

  • Altshuler GL, Dziewulski DM, Sowek JA and Belfort G (1986) Continuous hybridoma growth and monoclonal antibody production in hollow fiber reactors-separators. Biotechnol. Bioeng. 28: 646–658.

    Google Scholar 

  • Bergmeyer HU (ed) (1985) Methods of Enzymatic Analysis 8, VCH Verlagsgesellschaft Weinheim a) 357–363; b) 454–461.

    Google Scholar 

  • Blüml G, Reiter M, Zach N, Gaida T, Schmatz C, Strutzenberger C, Mohr T and Katinger H (1992) Development of a new type of macroporous carrier. In: Spier RE, Griffiths JB and MacDonald C (eds) Animal Cell Technology: Development, Processes and Products (pp. 501–504), Oxford, Butterworth-Heinemann.

    Google Scholar 

  • Boraston R, Thompson PW, Garland S and Birch JR (1984) Growth and oxygen requirements of antibody producing mouse hybridoma in suspension culture. Develop. Biol. Standard 55: 103–111.

    Google Scholar 

  • Cadic Ch, Dupuy B, Pianet I, Merle M, Margerin Ch and Bezian JH (1992) In vitro culture of hybridoma cells in agarose beads producing antibody secretion for two weeks. Biotechnol. Bioeng. 39: 108–112.

    Google Scholar 

  • Cooper PD, Burt AM and Wilson JN (1958) Critical effect of oxygen tension on rate of growth of animal cells in continuous suspension culture. Nature 182: 1508–1509.

    Google Scholar 

  • Flickinger M. (November 13–15, 1991) Proceedings of the 4th Annual Meeting of the Japanese Association for Animal Cell Technology. Fukukoka, Japan, to be published.

  • Frame KK and Hu WS (1991) Kinetic study of hybridoma cell growth in continuous culture: 1. A model for nonproducing cells. Biotechnol. Bioeng. 37: 55–64.

    Google Scholar 

  • Glacken MW, Fleischaker E and Sinskey AJ (1986) Reduction of waste product excretion via nutrient control: Possible strategies for maximising product and cell yields on serum in cultures of mammalian cells. Biotechnol. Bioeng. 28: 1376–1389.

    Google Scholar 

  • Hambach B, Biselli M, Runstadler PW and Wandrey C (1992) Development of a reactor integrated aeration system for cultivation of animal cells in fluidized beds. In: Spier RE, Griffiths JB and MacDonald C (eds) Animal Cell Technology: Development, Processes and Products (pp. 381–384). Oxford, Butterworth & Heinemann.

    Google Scholar 

  • Handa-Corrigan A, Nikolay S and Spier RE (1991) Biochemical control of monoclonal antibody secretion in hollow fibre bioreactors. In: Spier RE, Griffiths JB and Meignier B (eds) Production of Biologicals from Animal Cells in Culture (pp. 489–494), Oxford, Butterworth & Heinemann.

    Google Scholar 

  • Hayman EG, Ray NG and Runstadler PW (1988) In: Moody DU and Baker PE (eds) Bioreactors and Biotransformations (pp. 99–110), London and New York, Elsevier.

    Google Scholar 

  • Hiller GW, Aeschlimann AD, Clark DS and Blanch HW (1991) A kinetic analysis of hybridoma growth and metabolism in continuous suspension culture on serum-free medium. Biotechnol. Bioeng. 38: 733–741.

    Google Scholar 

  • Keller J, Dunn IJ and Heinzle J (1991) Improved performance of the fluidized bed reactor for the cultivation of animal cells. In: Spier, RE, Griffiths JB and Meignier B (eds) Production of Biologicals from Animal Cells in Culture (pp. 513–515), Oxford, Butterworth-Heinemann.

    Google Scholar 

  • Kratje RB and Wagner R (1992) Evaluation of production of recombinant human interleukin-2 in fluidized bed bioreactor. Biotechnol. Bioeng. 39: 233–242.

    Google Scholar 

  • Lazar A, Reuveny S, Mizrahi A, Avtalion M and Whiteside JP (1987) Production of biologicals by animal cells immobilized in a polyurethane foam matrix. In: Spier RE and Griffiths JB (eds) Modern Approaches to Animal Cell Technology (pp. 437–448), Guildford, Butterworth.

    Google Scholar 

  • Lee GM and Palsson BO (1990) Immobilization can improve the stability of hybridoma antibody productivity in serum-free media. Biotechnol. Bioeng. 36: 1049–1055.

    Google Scholar 

  • Lehmann J, Vorlop J and Büntemeyer H (1988) Bubble-free reactors and their development for continuous cell recycle. In: Spier RE, Griffiths JB (eds) Animal Cell Biotechnology, Vol 3 (pp. 222–237), London, Academic Press.

    Google Scholar 

  • Looby D and Griffiths JB (1988) Fixed bed porous glass sphere (porosphere) bioreactors for animal cells, Cytotechnology 1: 339–346.

    Google Scholar 

  • Lüllau E, Dreisbach C, Grogg AF, Biselli M and Wandrey C (1992) Immobilization of animal cells on chemically modified siran carriers. In: Spier RE, Griffiths JB and MacDonald C (eds) Animal Cell Technology: Development, Processes and Products (pp. 469–475), Oxford, Butterworth-Heinemann.

    Google Scholar 

  • Marc A, Wagner A, Martial A, Goergen JL, Engasser JM, Geaugey V and Pinton H (1991) Potential and pitfalls of using LDH release for the evaluation of animal cell death kinetics. In: Spier RE, Griffiths JB and Meignier B (eds) Production of Biologicals from Animal Cells in Culture (pp. 569–574), Oxford, Butterworth-Heinemann.

    Google Scholar 

  • Meilhoc E., Wittrup KD and Bailey JE (1990) Influence of dissolved oxygen concentration on growth, mitochondrial function and antibody production of hybridoma cells in batch culture. Bioproc. Eng. 5: 263–274.

    Google Scholar 

  • Miller WM, Blanch HW and Wike CR (1988a) A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: Effect of nutrient concentration, dilution rate and pH. Biotechnol. Bioeng. 32: 947–965.

    Google Scholar 

  • Miller WM, Wilke CR and Blanch HW (1988b) Transient responses of hybridoma metabolism to changes in the oxygen supply rate in continuous culture. Bioproc. Eng. 3: 103–111.

    Google Scholar 

  • Ozturk SS and Palsson BO (1990) Effect of dissolved oxygen on hybridoma cell growth metabolism and antibody production kinetics in continuous culture. Biotechnol. Progr. 6: 437–446.

    Google Scholar 

  • Ozturk SS and Palsson BO (1991a) Growth, metabolic and antibody production kinetics of hybridoma cell culture: 1. Analysis of data from controlled batch reactors. Biotechnol. Progr. 7: 471–480.

    Google Scholar 

  • Ozturk SS and Palsson BO (1991b) Growth, metabolic and antibody production kinetics of hybridoma cell culture: 2. Effect of serum concentration, dissolved oxygen concentration and medium pH in a batch reactor. Biotechnol. Progr. 7: 481–494.

    Google Scholar 

  • Phillips HA, Scharer JM, Bols NC and Moo-Young M (1987) Effect of oxygen on antibody productivity in hybridoma culture. Biotechnol. Lett. 9(11): 745–750.

    Google Scholar 

  • Pirt SJ (1965) The maintenance energy of bacteria in growing cultures. Proc. Royal. Soc. 163: 224–231.

    Google Scholar 

  • Ray NG, Karkare SB and Runstadler PW (1989) Cultivation of hybridoma cells in continuous culture: Kinetics of growth and product formation. Biotechnol. Bioeng. 33: 724–730.

    Google Scholar 

  • Reuveny S, Velez D, Miller L and Macmillan JD (1986) Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors. J. Immunol. Meth. 86: 61–69.

    Google Scholar 

  • Runstadler PW and Cernek R (1988) Large-scale fluidized-bed, immobilized cultivation of animal cells at high densities. In: Spier RE and Griffiths JB (eds) Animal Cell Biotechnology Vol. 3 (pp. 306–320), London, Academic Press.

    Google Scholar 

  • Runstadler PW, Ozturk SS and Ray NG. An evaluation of hybridoma cell specific productivity: Perfused immobilized, continuous suspension and batch suspension cultures, Proceedings of the 4th Annual Meeting of the Japanese Association for Animal Cell Technology. Fukukoka, Japan (1991), Kluwer Academic Publishers, Dordrecht, The Netherlands, to be published.

  • Wilson DW, Erecinska M, Drown C and Silver IA (1979) The oxygen dependency of cellular energy metabolism. Arch. Biochem. Biophys. 195(2): 485–493.

    Google Scholar 

  • Zielke HR, Zielke CL and Ozand PT (1984) Glutamine: A major energy source for cultured mammalian cells. Fed. Proc. 43: 121–125.

    Google Scholar 

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Author notes

  1. J. Thömmes

    Present address: Institut für Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, P.O. Box 2050, D-52404, Jülich, Germany

Authors and Affiliations

  1. Institut für Biotechnologie, Forschungszentrum Jülich, P.O. Box 1913, D-52425, Jülich

    J. Thömmes, J. Gätgens, M. Biselli & C. Wandrey

  2. VERAX Corp., 6 Etna Road, 03766, Lebanon, NH, USA

    P. W. Runstadler

Authors
  1. J. Thömmes
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  2. J. Gätgens
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  3. M. Biselli
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  4. P. W. Runstadler
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  5. C. Wandrey
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Thömmes, J., Gätgens, J., Biselli, M. et al. The influence of dissolved oxygen tension on the metabolic activity of an immobilized hybridoma population. Cytotechnology 13, 29–39 (1993). https://doi.org/10.1007/BF00749973

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  • DOI: https://doi.org/10.1007/BF00749973

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