Abstract
Inhibition caused by rapid changes in the environment has earlier been observed in hybridoma cultures following deliberate step-changes in the culture environment. This paper presents evidence of similar effects occurring during the normal span of continuous cultures fed enriched medium at low dilution rates (0.002–0.005 1/h). The effect of this observation on optimisation is discussed. In continuous culture at a dilution rate of 0.013 1/h, a viable cell density of 4×109 cells/l was achieved by gradually increasing the nutrient concentration in the feed medium. The MAb titre was 200 mg/l representing a 6-fold increase compared to batch culture and a 2-fold increase compared to continuous culture using standard medium.
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References
Dodge TC, Ji G and Hu W (1987) Loss of viability in hybridoma cell culture—a kinetic study. Enzyme Microb. Technol. 9: 607–611
Flickinger MC, Goebel NK and Bohn MA (1990) Determination of specific monoclonal antibody secretion rate during very slow hybridoma growth. Bioprocess Engineering 5: 155–164
Jones RP and Greenfield PF (1984) Kinetics of apparent cell death in yeasts induced by ethanol. Biotechnology Letters 6: 471–476
Jones RP and Greenfield PF (1985) Replicative inactivation and metabolite inhibition in yeast ethanol fermentation. Biotechnology Letters 7: 223–228
Luan YT, Mutharasan R and Magee WE (1987) Effect of various glucose/glutamine ratios on hybridoma growth, viability and monoclonal antibody formation. Biotechnology Letters 9: 535–538
Miller WM, Wilke CR and Blanch HW (1988a) Transient response of hybridoma cells to lactate and ammonia pulse and step changes in continuous culture. Bioprocess Engineering 3: 113–122
Miller WM, Blanch HW and Wilke CR (1988b) A kinetic analysis of hybridoma growth and metabolism in batch and continuous culture: Effect of nutrient concentration, dilution rate and pH. Biotechnology and Bioengineering 32: 947–965
Newland M (1990) Growth limitation of hybridoma cells by ammonia. Masters Thesis (University of Queensland, Brisbane, Australia)
Nielsen LK (1989) Modelling growth of and antibody production by hybridomas using an unstructured model. Masters Thesis (DtH, Lyngby, Denmark).
Nielsen LK, Niloperbowo W, Reid S and Greenfield PF (1991a) Modelling growth of and antibody production by hybridomas in glutamine limited suspension cultures. In: Spier R, Griffiths JB & Meinier B (eds.) Production of Biologicals from Animal Cells in Culture (10th meeting of ESACT, Avignon, France, 1990). Butterworth & Heinemann, Oxford, UK, pp. 625–630
Nielsen LK, Smyth SK and Greenfield PF (1991b) Haemocytometer Cell Count Distributions: Implications of Non-Poisson Behaviour. Biotechnology Progress 7: 560–563
Oyaas K, Berg TM, Bakke O and Levine DW (1989) Hybridoma growth and antibody production under conditions of hyperosmotic stress. In: Spier R, Griffiths JB, Stephenne J & Crooy PJ (eds.) Advances in Animal Cell Biology and Technology for Bioprocesses (9th meeting of ESACT, Knokke-Heist, Belgium, 1988). Butterworth & Co., Kent, UK, pp. 212–218
Ray NG, Karkare SB and Runstadler PW, Jr. (1989) Cultivation of hybridoma cells in continuous culture: Kinetics of growth and product formation. Biotechnology and Bioengineering 33: 724–730
Reuveny S, Velez D, Macmillan JD and Miller L (1986) Journal of Immunological Methods 86: 53–59
Robinson DK and Memmert KW (1991) Kinetics of Recombinant Immunoglobulin Production by Mammalian Cells in Continuous Culture. Biotechnology and Bioengineering 38: 972–976
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Nielsen, L.K., Niloperbowo, W., Reid, S. et al. Avoiding rapid growth at high cell densities: A potentially important optimisation criterion for hybridoma cultures. Cytotechnology 9, 21–27 (1992). https://doi.org/10.1007/BF02521728
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DOI: https://doi.org/10.1007/BF02521728