Abstract
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.
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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.
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.
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.
Dalili M, Sayles GD and Ollis DF (1990) Glutamine-limited batch hybridoma growth and antibody production: experiment and model. Biotechnol. Bioeng. 36: 74–82.
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.
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.
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.
Hayter PM, Kirby NF and Spier RE (1992) Relationship between hybridoma growth and monoclonal antibody production. Enzyme Microb. Technol. 14: 454–461.
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.
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.
Kerr MA (1990) The structure and function of human IgA. Biochem. J. 271: 285–296.
Lindh E (1975) Increased resistance of immunoglobulin A dimers to proteolytic degradation after binding of secretory component. J. Immunol. 114: 284–286.
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.
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.
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.
Miller CJ, McGhee J and Gardner MB (1992) Biology of disease: mucosal immunity, HIV transmission, and AIDS. Lab. Investigation 68 (2): 129–145.
Newland M, Greenfield PF and Reid S (1990) Hybridoma growth limitations: the roles of energy metabolism and ammonia production. Cytotechnology 3: 215–229.
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.
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.
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.
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.
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.
Tritsch GL and Moore GE (1962) Spontaneous decomposition of glutamine in cell culture media. Exp. Cell Res. 28: 360–364.
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.
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Stoll, T., Perregaux, C., von Stockar, U. et al. Production of immunoglobulin A in different reactor configurations. Cytotechnology 17, 53–63 (1995). https://doi.org/10.1007/BF00749221
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DOI: https://doi.org/10.1007/BF00749221