Design and performance of a trickle-bed bioreactor with immobilized hybridoma cells
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A trickle-bed system employing inert matrices of vermiculite or polyurethane foam packed in the downcomer section of a split-flow air-lift reactor has been developed for hybridoma culture to enhance antibody productivity. This quiescent condition favoured occlusion and allowed the cells to achieve densities twelve fold greater (12.8×106 cells/ml reactor for polyurethane foam) than in free cell suspension.
The reactor was operated in a cyclic batch mode whereby defined volumes of medium were periodically withdrawn and replaced with equal volumes of fresh medium. The pH of the medium was used as the indicator of the feeding schedule. Glucose, lactate and ammonia concentrations reached a stationary value after 5 days. With vermiculite packing, a monoclonal antibody (MAb) concentration of 2.4 mg/l was achieved after 12 days. The MAb concentration declined then increased to a value of 1.8 mg/l. In the polyurethane foam average monoclonal antibody (MAb) concentrations reached a stationary value of 1.1 mg/l in the first 20 days and increased to a new stationary state value of 2.1 mg/l for the remainder of the production. MAb productivity in the trickle-bed reactor was 0.3 mg/l·d (polyurethane foam) and 0.18 mg/l.d (vermiculite) in comparison to 0.12 mg/l·d for free cell suspension. This trickle-bed system seems to be an attractive way of increasing MAb productivity in culture.
Key wordshybridoma immobilization tricle-bed polyurethane foam vermiculite MAb productivity
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- Buehler RJ (1984) Production of monoclonal antibody from gel entrapped hybridoma cells. Karyon Technol News 1 (1): 3.Google Scholar
- Dean RC Jr, Karkare SB, Phillips PG, Ray NG and Runstadler PW (1986) Continuous cell culture with fluidized sponge beads for large scale production of medical proteins. Verax Corporation Report, New Hampshire, USA.Google Scholar
- Handa-Corrigan A, Emery AN and Spier RE (1987) On the evaluation of gas liquid interfacial effects on hybridoma viability in bubble column reactors. Dev Biol Stand 66: 241–253.Google Scholar
- Karkare SB, Phillips PG, Burke DH and Dean RC (1985) Continuous production of monoclonal antibodies by chemostatic and immobilized hybridoma culture. In: Feder J and Tolbert WR (eds) Large Scale Mammalian Cell Culture, Academic Press, New York.Google Scholar
- Lazar A (1988) Immobilized animal cell culture systems for production of biologicals. Presented at ACS Conference, Boston, USA.Google Scholar
- Mavituna F, Wilkinson AK, Williams PD and Park JM (1987) Production of secondary metabolites by immobilized plant cells in novel bioreactors. Presented at the International Conference on Bioreactors and Biotransformations, Scotland.Google Scholar
- Murdin AD, Thorpe JS, Kirby N, Groves DE and Spier RE (1987) Immobilization and growth of hybridomas in packed beds. Presented at the International Conference on Bioreactors and Biotransformations, Scotland.Google Scholar
- Nilsson K, Scheirer W, Katinger HWD, Mosbach K (1986b) Production of monoclonal antibodies by agarose-entrapped hybridoma cells. In: Mosbach K (ed) Academic Press, London, Meth Enzymol 121: 352–360.Google Scholar
- Nilsson K (1987) Mammalian cell culture. In: Mosbach K (ed) Academic Press, London, Meth Enzymol 135: 183–193.Google Scholar
- Phillips HA (1991) Evaluation of hybridoma culture systems. PhD Thesis. University of Waterloo, Waterloo, Ontario, Canada.Google Scholar
- Phillips J (1973) Dye-exclusion tests for viability. In: Kruse PF Jr and Patterson MR Jr (ed) Tissue Culture Methods of Applications. Academic Press, New York.Google Scholar