Skip to main content
Log in

Hybridomas in a bioreactor cascade: modeling and determination of growth and death kinetics

  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Hybridomas were cultured under steady-state conditions in a series of two continuous stirred-tank reactors (CSTRs), using a serum-free medium. The substrate not completely converted in the first CSTR, was transported with the cells to the second one and very low growth rates, high death rates, and lysis of viable cells were observed in this second CSTR. These conditions are hardly accessible in a single vessel, because such experiments would be extremely time-consuming and unstable due to a low viability. In contrast to what is often observed in literature, kinetic parameters could thus be derived without the neccessity for extrapolation to lower growth rates. Good agreement with literature averages for other hybridomas was found. Furthermore, showing that the reactor series is a valuable research tool for kinetic studies under extreme conditions, the possibility to observe cell death under stable and defined steady-state conditions offers interesting opportunities to investigate apoptosis and necrosis. Additionally, a model was developed that describes hybridoma growth and monoclonal antibody production in the bioreactor cascade on the basis of glutamine metabolism. Good agreement between the model and the experiments was found.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

MAb:

Monoclonal antibody

References

  • Aeschlimann A, Di Stasi L and von Stockar U (1990) Continuous production of lactic acid from whey permeate by Lactobacillus helveticus in two chemostats in series. Enzyme Microb. Technol. 12: 926–932.

    Google Scholar 

  • Al-Rubeai M, Emery AN, Chalder S and Jan DC (1992) Specific monoclonal antibody productivity and the cell cycle-comparisons of batch, continuous and perfusion cultures. Cytotechnology 9: 85–97.

    Google Scholar 

  • Button DK (1985) Kinetics of nutrient-limited transport and microbial growth. Microbiol. Rev. 49: 270–297.

    Google Scholar 

  • De Gooijer CD, Bakker WAM, Beeftink HH and Tramper J (1996) Bioreactors in series: an overview of design procedures and practical applications. Enzyme Microb. Technol. 18: 202–219.

    Google Scholar 

  • Frame KK and Hu W-S (1991a) Kinetic study of hybridoma cell growth in continuous culture. I. A model for non-producing cells. Biotechnol. Bioeng. 37: 55–64.

    Google Scholar 

  • Frame KK and Hu W-S (1991b) Kinetic study of hybridoma cell growth in continuous culture: II. Behavior of producers and comparison to nonproducers. Biotechnol. Bioeng. 38: 1020–1028.

    Google Scholar 

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

    Google Scholar 

  • Goergen JL, Marc A and Engasser JM (1993) Determination of cell lysis and death kinetics in continuous hybridoma cultures from the measurement of lactate dehydrogenase release. Cytotechnology 11: 189–195.

    Google Scholar 

  • Hill GA and Robinson CW (1989) Minimum tank volumes for CFST bioreactors in series. Can. J. Chem. Eng. 67: 818–824.

    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 

  • Linardos TI, Kalogerakis N, Behie LA and Lamontagne LR (1991) The effect of specific growth rate and death rate on monoclonal antibody production in hybridoma chemostat cultures. Can. J. Chem. Eng. 69: 429–438.

    Google Scholar 

  • Lüdemann I, Pörtner R and Märkl M (1994) Effect of NH3 on the cell growth of a hybridoma cell line. Cytotechnology 14: 11–20.

    Google Scholar 

  • Martens DE, Sipkema EM, De Gooijer CD, Beuvery EC and Tramper J (1995) A combined cell-cycle and metabolic model for the growth of hybridoma cells in steady-state continuous culture. Biotechnol. Bioeng. 48: 49–65.

    Google Scholar 

  • Mercille S and Massie B (1994) Induction of apoptosis in nutrient-deprived cultures of hybridoma and myeloma cells. Biotechnol. Bioeng. 44: 1140–1154.

    Google Scholar 

  • Miller WM, Blanch HW and Wilke CR (1988) 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 

  • Ozturk SS and Palsson BO (1990) Chemical decomposition of glutamine in cell culture media: effect of media type, pH and serum concentration. Biotechnol. Prog. 6: 121–128.

    Google Scholar 

  • Ozturk SS, Riley MR and Palsson BO (1992) Effects of ammonia and lactate on hybridoma growth, metabolism, and antibody production. Biotechnol. Bioeng. 39: 418–431.

    Google Scholar 

  • Pirt SJ (1975) Principles of microbe and cell cultivation. Blackwell, Oxford.

    Google Scholar 

  • Press WH, Flannery BP, Teukolsky SA and Vetterling WA (1989) Numerical recipes in Pascal. Cambridge university press, Cambridge.

    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 

  • Shama G (1988) Developments in bioreactors for fuel ethanol production. Process Biochem. 10: 138–145.

    Google Scholar 

  • Shimizu K and Matsubara M (1987) Product formation patterns and the performance improvement for multistage continuous stirred tank fermentors. Chem. Eng. Comm. 52: 61–74.

    Google Scholar 

  • Singh RP, Al-Rubeai M, Gregory CD and Emery AN (1994) Cell death in bioreactors: a role for apoptosis. Biotechnol. Bioeng. 44: 720–726.

    Google Scholar 

  • Truskey GA, Nicolakis DP, DiMasi D, Haberman A and Swartz RW (1990) Kinetic studies and unstructured models of lymphocyte metabolism in fed-batch culture. Biotechnol. Bioeng. 36: 797–807.

    Google Scholar 

  • Van der Pol L, Bakker WAM and Tramper J (1992) Effect of low serum concentrations (0%–2.5%) on growth, production, and shear sensitivity of hybridoma cells. biotechnol. Bioeng. 40: 179–182.

    Google Scholar 

  • Van der Pol L, Zijlstra G, Thalen M and Tramper J (1990) Effect of serum concentration on production of monoclonal antibodies and on shear sensitivity of a hybridoma. Bioproc. Eng. 5: 241–245.

    Google Scholar 

  • Van Oers JWAM, Tilders FJH and Berkenbosch F (1989) Characterization of a rat monoclonal to rat/human corticotropin releasing factor. Endocrinology 124: 1239–1246.

    Google Scholar 

  • Venables DC, Boraston RC and Bushell ME (1993) Two-stage chemostat studies of hybridoma growth, nutrient utilisation, and monoclonal antibody production. In: S. Kaminogawa et al. (eds.), Animal cell technology, basic and applied aspects. Vol. 5 (pp. 585–594) Kluwer academic publishers, The Netherlands.

    Google Scholar 

  • Zwietering MH, Jongenburger I, Rombouts FM and Van't Riet K (1990) Modeling of the bacterial growth curve. Appl. Environ. Microbiol. 56: 1875–1881.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Nomenclature

C AConcentration of any (mol m-3) component A

D Dilution rate (s-1)

K dDeath-rate constant (mol m-3)

K lLysis-rate constant (mol m-3)

K sMonod constant (mol m-3)

m Maintenance coefficient (mol cell-1 s-1)

q Specific consumption (mol cell-1 s-1) or production rate

t Time (s)

X Cell concentration (cell m-3)

Y Yield coefficient (cell mol-1)

Greek symbols

μ d Specific death rate (s-1)

μ l Specific lysis rate (s-1) of viable cells

μ net Net specific growth (s-1) rate

μ true True specific growth (s-1) rate

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bakker, W.A.M., Schäfer, T., Beeftink, H.H. et al. Hybridomas in a bioreactor cascade: modeling and determination of growth and death kinetics. Cytotechnology 21, 263–277 (1996). https://doi.org/10.1007/BF00365349

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00365349

Key words

Navigation