A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: Effect of nutrient concentration, dilution rate, and pH

Biotechnology and Bioengineering - Tập 32 Số 8 - Trang 947-965 - 1988
William M. Miller1, Harvey W. Blanch1, C. R. Wilke1
1Department of Chemical Engineering, University of California, Berkeley, California 94720

Tóm tắt

AbstractHybridomas are finding increased use for the production of a wide variety of monoclonal antibodies. Understanding the roles of physiological and environmental factors on the growth and metabolism of mammalian cells is a prerequisite for the development of rational scale‐up procedures. An SP2/0‐derived mouse hybridoma has been employed in the present work as a model system for hybridoma suspension culture. In preliminary shake flask studies to determine the effect of glucose and glutamine, it was found that the specific growth rate, the glucose and glutamine metabolic quotients, and the cumulative specific antibody production rate were independent of glucose concentration over the range commonly employed in cell cultures. Only the specific rate of glutamine uptake was found to depend on glutamine concentration. The cells were grown in continuous culture at constant pH and oxygen concentration at a variety of dilution rates. Specific substrate consumption rates and product formation rates were determined from the steady state concentrations. The specific glucose uptake rate deviated from the maintenance energy model1at low specific growth rates, probably due to changes in the metabolic pathways of the cells. Antibody production was not growth‐associated; and higher specific antibody production rates were obtained at lower specific growth rates. The effect of pH on the metabolic quotients was also determined. An optimum in viable cell concentration was obtained between pH 7.1 and 7.4. The viable cell number and viability decreased dramatically at pH 6.8. At pH 7.7 the viable cell concentration initially decreased, but then recovered to values typical of pH 7.1–7.4. Higher specific nutrient consumption rates were found at the extreme pH values; however, glucose consumption was inhibited at low pH. The pH history also influenced the behavior at a given pH. Higher antibody metabolic quotients were obtained at the extreme pH values. Together with the effect of specific growth rate, this suggests higher antibody production under environmental or nutritional stress.

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Tài liệu tham khảo

Pirt S. J., 1975, Principles of Microbe and Cell Cultivation

Hubbard R., 1983, Topics in Enzyme and Fermentation Biotechnology, 196

10.1038/314446a0

10.1093/nar/12.9.3791

Colby C. B., 1984, J. Immunol., 133, 3091, 10.4049/jimmunol.133.6.3091

Wilson T., 1984, Biol Technology, 2, 753

Spier R. E., 1982, J. Chem. Technol. Biotechnol., 23, 304, 10.1002/jctb.5030320134

10.1016/0167-7799(83)90032-X

10.1002/btpr.5420010404

10.1016/0168-1656(85)90028-8

10.1002/abio.370020102

Mizrahi A., 1984, Dev. Biol. Standard., 55, 93

10.1016/0022-1759(86)90264-4

10.1016/0022-1759(86)90265-6

Lavery M., 1985, Dev. Biol. Standard., 60, 199

van Wezel A. L., 1985, Dev. Biol. Standard., 60, 229

10.1016/0022-1759(83)90070-4

J. R.Birch P. W.Thompson K.Lambert andR.Boraston “The Large Scale Cultivation of Hybridoma Cells Producing Monoclonal Antibodies ” ACS National Meeting Philadelphia PA August 27 1984.

Boraston R., 1984, Dev. Biol. Standard., 55, 103

10.1007/BF02135954

10.1007/BF02615311

10.1016/0014-4827(76)90387-6

10.1016/S0021-9258(17)30124-2

Zielke H. R., 1984, Fed. Proc. (FASEB), 43, 121

10.1016/0168-1656(84)90004-X

M. W.GlackenandA. J.Sinskey “Modelling and Optimization of Fed‐Batch Cultures of Hybridoma Cells.” AIChE National Meeting Chicago IL November 11 1985.

10.1084/jem.161.1.53

10.1007/BF01025202

10.1063/1.432215

10.1002/bit.260200114

Low K., 1985, Dev. Biol. Standard., 60, 73

10.1002/jcp.1040950106

Waymouth C., 1981, The Growth Requirements of Vertebrate Cells in Vitro, 103

10.1002/jcp.1041040316

Wice B. M., 1981, J. Biol. Chem., 256, 7812, 10.1016/S0021-9258(18)43351-0

10.1016/0304-4165(81)90242-7

10.1098/rspb.1967.0073

10.1002/jcp.1040890105

10.1007/BF01027808

10.1002/bit.260100604

10.1007/BF00499486

Low K., 1985, Dev. Biol. Standard., 60, 17

Schlesinger M. J., 1982, Heat Shock, from Bacteria to Man

Demain A. L., 1968, Lloydia, 31, 395

Merten O‐W., 1985, Dev. Biol. Standard., 60, 219

10.1002/jcp.1040820102

10.1002/bit.260130403

Birch J. R., 1980, Dev. Biol. Standard., 46, 59

10.1016/0014-4827(63)90308-2

W. M.Miller H. W.Blanch andC. R.Wilke “Kinetic Analysis of Hybridoma Growth in Continuous Suspension Culture ” ACS National Meeting Anaheim CA

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Biotechnology and Bioengineering

Tập 32 Số 8

947-965

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