, Volume 17, Issue 1, pp 53–63 | Cite as

Production of immunoglobulin A in different reactor configurations

  • T. Stoll
  • C. Perregaux
  • U. von Stockar
  • I. W. Marison
Technical Report


A murine hybridoma line (Zac3), secreting an IgA monoclonal antibody, was cultivated in different systems: a BALB/c mouse, a T-flask, a stirred-tank bioreactor and a hollow fiber reactor. These systems were characterized in terms of cell metabolism and performances for IgA production. Cultures in T-flask and batch bioreactor were found to be glutamine-limited. Ammonia and lactate were produced in significant amounts. IgA productivity was found to be constant and growth associated. Final IgA concentration was similar in both systems. In fed-batch cultures, supplemented with glutamine and glucose, maximum viable cell concentration was increased by 60% and final IgA concentration by 155%. The hollow fiber reactor was able to produce very large amounts of IgA at very high concentrations, similar to the value found in ascites fluid. The productivity ofZac3 is similar to the values reported for IgG-producing cell lines.

Key words

Hollow fiber reactor hybridoma cells immunoglobulin A reactor comparison 


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  1. Amerongen HM, Michetti P, Weltzin R, Lee TH, Kraehenbuhl J-P and Neutra MR (1991) Transepithelial delivery of a recombinant HIV protein on hydroxyapatite for production of monoclonal anti-gp 120 IgA antibodies. J. Cell Biol. 115: 237a.Google Scholar
  2. Apter FM, Lencer WI, Mekalanos JJ and Neutra MR (1991) Analysis of epithelial protection by monoclonal IgA antibodies directed against cholera toxin B subunit. J. Cell Biol. 115: 399A.Google Scholar
  3. Chandler JP (1987) Factors influencing monoclonal antibody production in mouse ascites fluid. In: Seaver SS (ed.) Commercial Production of Monoclonal Antibodies: A Guide for Scale-Up (pp. 75–92). Marcel Dekker Inc., New York.Google Scholar
  4. Dalili M, Sayles GD and Ollis DF (1990) Glutamine-limited batch hybridoma growth and antibody production: experiment and model. Biotechnol. Bioeng. 36: 74–82.Google Scholar
  5. Dhainaut F, Meterreau JL, Mas MP, Potentini C and Mignot G (1992) Scale-up of human IgG1 production by a lymphoblastoid cell line in hollow fiber systems. In: Spier RE (ed.) Animal Cell Technology: Developments, Processes & Products ESACT 11th Meeting (pp. 527–529). Butterworth-Heinemann, Oxford.Google Scholar
  6. 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
  7. Hassel T, Gleave S and Butler M (1991) Growth inhibition in animal cell culture: the effect of lactate and ammonia. Appl. Biochem. Biotechnol. 30: 29–41.Google Scholar
  8. Hayter PM, Kirby NF and Spier RE (1992) Relationship between hybridoma growth and monoclonal antibody production. Enzyme Microb. Technol. 14: 454–461.Google Scholar
  9. Jöbses I, van Zutphen P, Oomens J, van Os A and Schönherr O (1992) Scaling-up of a hollow fibre reactor for animal cell cultivation. In: Spier RE (ed.) Animal Cell Technology: Developments. Processes & Products, ESACT 11th Meeting (pp. 517–523). Butterworth-Heinemann, Oxford.Google Scholar
  10. Keren DF and Silbart LK (1992) Strategies to achieve mucosal immunity. In: Isaacson R (ed.) Recombinant DNA Vaccines: Rationale and Method (pp. 145–168). Marcel Dekker Inc., New York.Google Scholar
  11. Kerr MA (1990) The structure and function of human IgA. Biochem. J. 271: 285–296.Google Scholar
  12. Lindh E (1975) Increased resistance of immunoglobulin A dimers to proteolytic degradation after binding of secretory component. J. Immunol. 114: 284–286.Google Scholar
  13. Mestecky J and McGhee JR (1987) Immunoglobulin A (IgA): molecular and cellular interactions involved in IgA biosynthesis and immune response. Adv. Immunol. 40: 153–245.Google Scholar
  14. Michetti P, Mahan MJ, Slauch JM, Mekalanos JJ and Neutra MR (1992) Monoclonal secretory immunoglobulin A protects mice against oral challenge with invasive pathogenSalmonella typhimurium. Infect. Immun. 60 (5): 1786–1792.Google Scholar
  15. Michetti P, Perregaux C, Amerongen HM, Neutra MR, Ada-Ochea H and Kraehenbuhl J-P (1994) Mucosal immune protection against retroviral infection: the mouse mammary tumor virus model. Nature, submitted.Google Scholar
  16. Miller CJ, McGhee J and Gardner MB (1992) Biology of disease: mucosal immunity, HIV transmission, and AIDS. Lab. Investigation 68 (2): 129–145.Google Scholar
  17. Newland M, Greenfield PF and Reid S (1990) Hybridoma growth limitations: the roles of energy metabolism and ammonia production. Cytotechnology 3: 215–229.Google Scholar
  18. Offit PA and Clark HF (1985) Protection against rotavirus-induced gastroenteritis in a murine model by passively acquired gastroin-testinal but not circulating antibodies. J. Virol. 54, 1: 58–64.Google Scholar
  19. Ozturk SS and Palsson BO (1991) Growth, metabolic, and antibody kinetics of hybridoma cell culture: 1. Analysis of data from controlled batch reactors. Biotechnol. Prog. 7: 471–480.Google Scholar
  20. Reitzer LJ, Wice BM and Kennel D (1979) Evidence that glutamine, not sugar, is the major energy source for cultured HeLa cells. J. Biol. Chem. 254 (8): 2669–2676.Google Scholar
  21. Schneider M, Marison IW and von Stockar U (1993) Hybridoma cell culture in a membrane reactor within situ ammonia removal. Poster presented at the 6th European Congress on Biotechnology, Firenze, 13–17 June 1993.Google Scholar
  22. Stoll T, Pugeaud P, von Stockar U and Marison IW (1994) A simple HPLC technique for accurate monitoring of mammalian cell metabolism. Cytotechnology 14: 123–128.Google Scholar
  23. Tritsch GL and Moore GE (1962) Spontaneous decomposition of glutamine in cell culture media. Exp. Cell Res. 28: 360–364.Google Scholar
  24. Winner L III, Mack J, Weltzin R, Mekalanos JJ, Kraehenbuhl J-P and Neutra MR (1991) New model for analysis of mucosal immunity: intestinal secretion of specific monoclonal immunoglobulin A from hybridoma tumors protects againstVibrio cholerae infection. Infect. Immun. 59 (3): 977–982.Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • T. Stoll
    • 1
  • C. Perregaux
    • 1
  • U. von Stockar
    • 1
  • I. W. Marison
    • 1
  1. 1.Institute of Chemical EngineeringSwiss Federal Institute of Technology (EPFL)LausanneSwizerland

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