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. 1997 May;24(1):47–54. doi: 10.1023/A:1007914004727

Optimization of monoclonal antibody production: combined effects of potassium acetate and perfusion in a stirred tank bioreactor

Wangfun Fong 1,✉, Yuanxing Zhang 2, Pingpei Yung 1
PMCID: PMC3449610  PMID: 22358596

Abstract

To increase the yield of monoclonal antibody in a hybridoma culture, it is important to optimize the combination of several factors including cell density, antibody productivity per cell, and the duration of the culture. Potassium acetate enhances the production of antibodies by cells but sometimes depresses cell density. The production of anti-(human B-type red blood cell surface antigen) antibody by Cp9B hybridoma was studied. In batch cultures, potassium acetate inhibited Cp9B cells growth and decreased the maximal cell density but the productivity of antibody per cell was increased. The balance of the two effects resulted in a slight decline of antibody production. In a stirred tank bioreactor, the inhibitory effect of potassium acetate on cell density was overcome by applying the perfusion technique with the attachment of a cell-recycling apparatus to the bioreactor. In such a reactor, potassium acetate at 1 g l-1 did not cause a decrease in the cell density, and the antibody concentration in the culture supernatant was increased from 28 μg ml-1 to 38 μg ml-1. Potassium acetate also suppressed the consumption of glucose and the accumulation of lactate in batch cultures, but the glucose and lactate levels were kept stable by applying the perfusion technique in the stirred tank bioreactor.

Keywords: hybridoma, monoclonal antibody, stirred tank perfusion culture, potassium acetate

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References

  1. Al-Rubeai M, Emery AW. Mechanisms and kinetics of monoclonal antibody synthesis and secretion in synchronous and asynchronous hybridoma cell cultures. J, Biotechnol. 1990;16:67–86. doi: 10.1016/0168-1656(90)90066-K. [DOI] [PubMed] [Google Scholar]
  2. Al-Rubeai M, Emery AW, Chalder S, Han DC. Specific monoclonal antibody productivity and the cell cycle comparisons of batch, continuous and perfusion cultures. Cytotechnology. 1992;9:85–97. doi: 10.1007/BF02521735. [DOI] [PubMed] [Google Scholar]
  3. Fawcett J, Scott J. A rapid and precise method for the determination of urea. J. Clin. Path. 1960;13:126159. doi: 10.1136/jcp.13.2.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gaertner HG, Dhurjati P. Fractional factorial study of hybridomas behavior. 1. Kinetics of growth and antibody production. Biotechnol. Prog. 1993;9:298–308. doi: 10.1021/bp00021a009. [DOI] [PubMed] [Google Scholar]
  5. Gentry A, Morse PA, Potter VR. Pyrimidine metabolism in tissue culture cells derived from rat hepatomas. III. Relationship of thymidine to the metabolism of other pyrimidine nucleosides in suspension cultures derived from the Novikoff hepatoma. Cancer Res. 1965;25:517–525. [PubMed] [Google Scholar]
  6. Glacken MW, Adema E, Sinskey AJ. Mathematical descriptions of hybridoma culture kinetics: 1. Initial metabolic rates. Biotechnol. Bioeng. 1988;32:491–506. doi: 10.1002/bit.260320412. [DOI] [PubMed] [Google Scholar]
  7. Martinelle K and Häggström L (1994) Detoxification of ammonium in bioreactor cultivations. In: Animal Cell Technology: Products of Today, Prospects for Tomorrow, Spier RE, Griffiths JB, Berthold W (eds.), pp. 158–160, Butterworth-Heinemann.
  8. Merten OW. Batch production and growth kinetics of hybridomas. Cytotechnology. 1988;1:113–122. doi: 10.1007/BF00146811. [DOI] [PubMed] [Google Scholar]
  9. Miller WM, Blach HW, Wilke CR. A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: Effect of nutrient concentration, dilution rate, and pH. Biotechnol. Bioeng. 1988;32:947–965. doi: 10.1002/bit.260320803. [DOI] [PubMed] [Google Scholar]
  10. Oh SKW, Vig P, Chua F, Teo WK, Yap MGS. Substantial overproduction of antibodies by applying osmotic pressure and sodium butyrate. Biotechnol. Bioeng. 1993;42:601–620. doi: 10.1002/bit.260420508. [DOI] [PubMed] [Google Scholar]
  11. Oyaas K, Berg TM, Bakke O, Levine DW. Hybridoma growth and antibody production under conditions of hyperosmotic stress. In: Spier RE, Griffiths IB, Crooy PJ, editors. Advances in animal cell biology and technology for bioprocesses. Essex: Butterworth.; 1989. pp. 212–220. [Google Scholar]
  12. Oztark SS, Palsson BO. Effect of medium osmolarity on hybridoma growth, metabolism, and antibody production. Biotechnol. Bioeng. 1991;37:989–993. doi: 10.1002/bit.260371015. [DOI] [PubMed] [Google Scholar]
  13. Somenshein GE, Brawerman G. Differential translation of mouse myeloma messenger RNAs in a wheat germ cell-free system. Biochemistry. 1976;15:5501–5506. doi: 10.1021/bi00670a013. [DOI] [PubMed] [Google Scholar]
  14. Suzuki E, Ollis DF. Enhanced antibody production at slowed growth rates: Experimental demonstration and a structured model. Biotechnol. Prog. 1990;6:231–236. doi: 10.1021/bp00003a013. [DOI] [PubMed] [Google Scholar]
  15. Vits H, Hu W-S. Fluctuations in continuous mammalian cell bioreactors with retention. Biotechnol. Prog. 1992;8:397–403. doi: 10.1021/bp00017a004. [DOI] [PubMed] [Google Scholar]
  16. Xu D, Zhou Y, Zhang Y. Effects of potassium acetate on WuT3 cell growth and monoclonal antibody production. In: Two WK, Yap MGS, SKW, editors. Better living through innovative biochemical engineering. Singapore: Continental Press; 1994. pp. 226–228. [Google Scholar]

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