Abstract
Hybridoma cell growth and monoclonal antibody production were investigated with a laboratory-made system in which cells were grown in dialysis tubing (MW cut-off 25 kD). The dialysis system contained 10 ml of cell suspension and was immersed in 200 ml of culture medium which when replaced or was at 4-day intervals. With this system, monoclonal antibody concentrations similar to those observed in ascites (concentrations in the order of one gramme per liter) were obtained. With no medium replacement, the antibody production was 3.3 g/l and the cell productivity 3.2×10−8 μg of IgM produced per cell in one minute. With medium replacement the antibody production was higher, 4.4 g/l but the cell productivity was lower, 1.49×10−8 μg per cell in one minute. Cells cultivated in non-optimized conditions were better producers than cells growing in a good environment.
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Abbreviations
- FCS:
-
fetal calf serum
- Ig:
-
immunoglobulin
- MAb:
-
monoclonal antibody
- MW:
-
molecular weight
- MWCO:
-
molecular weight cut off
- RM:
-
replaced medium
- NRM:
-
non replaced medium
Literature
Adamson SR, Fritzpatrick SL, Behie LA, Gaucher GM and Lesser BH (1983)In vitro production of high titre monoclonal antibody by hybridoma cells in dialysis culture. Biotechnol. Lett. 5: 573–578.
Clark JTM (1987) Culture des cellules en masse: un examen des systemes actuellement disponibles pour la culture à grande échelle des cellules en suspension et des cellules adhérentes. Bio-Sciences 6: 51–53.
Comer MJ, Kearns M, Wahl J, Munster M, Lorentz T, Szperalski B, Koch S, Behrendt U and Brunner H (1990) Industrial production of monoclonal antibodies and therapeutic proteins by dialysis fermentation. Cytotechnology 3: 295–299.
Duval D, Demangel C, Munier-Johain K, Miossec S and Geahel I (1991) Factors controlling cell proliferation and antibody production in mouse hybridoma cells: I Influence of the amino acid supply. Biotechnol. Bioeng. 38: 561–570.
Eyl V, Mathiot B, Dusch M, Maugras M and Stoltz JF (1991) Comparaison de différentes méthodes de production d'anticorps monoclonaux. Inov. Tech. Biol. Med. 12: 457–466.
Glacken MW, Fleischaker RJ and Sinskey AJ (1985) Largescale production of mammalian cells and their products: engineering principles and barriers to scale up. Ann. N.Y. Acad. Sci. 413: 355–372.
Glassy MC, Tharakan JP and Chau PC (1988) Serum free media in hybridoma culture and monoclonal antibody production. Biotechnol. Bioeng. 32: 1015–1028.
Graf H, Rabaud JN and Egly JM (1991) Screening of various mammalian cell culture media to establish a downstream purification scheme. J. Immunol. Method. 139: 135–144.
Griffiths JB (1992) Animal cell culture processes — batch or continuous? J. Biotechnol. 22: 21–30.
Käsehagen C, Linz F, Kretzmer G, Scheper T, Schügerl K (1991) Enzyme Microb. Technol. 13: 873–881.
Lind W, Jäger V, Lucki-Lange M and Wagner R (1991) Characterization of protease activity in serum-free culture supernatants of hybridomas and recombinant mammalian cells. In: RE Spier, JB Griffiths, B Meignier (eds) 10th ESACT Meeting: Production of biologicals from animal cells in culture (pp. 196–202). Butterworth-Heinemann Ltd., Publishers, Oxford.
Kearns M (1990) Integrated design for mammalian cell culture. Bio/Technol. 8: 409–413.
Linardos TI, Kalogerakis N, Behie LA and Lamontagne LR (1992) Monoclonal antibody production in dialysed continuous suspension culture. Biotechnol. Bioeng. 39: 504–510.
Long WJ, Palombo A, Schofield TL and Emini EA (1988) Effects of culture media on murine hybridomas: definition of optimal conditions for hybridoma viability, cellular proliferation and antibody production. Hybridoma 7: 69–77.
Mancini G, Carbonara AO and Heremans JF (1965) Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry 2: 235–254.
Märkl H, Lechner M, Gotz F (1990) A new dialysis fermentor for the production of high concentrations of extracellular enzymes. J. Ferment. Bioeng. 69: 244–249.
Marquis CP, Harbour C, Barford JP and Low KS (1990) A comparison of different culture methods for hybridoma propagation and monoclonal antibody production. Cytotechnology 4: 69–76.
Mathiot B (1992) Etudes de méthodes de culture et de milieux contenant des protéines humaines pour la production d'anticorps monoclonaux. Thèse de Génie Biologique. Université de Nancy I.
Merten OW, Keller H, Siami K, Cabanie L and Leno M (1990) Batch production kinetics of hybridomas: pulse experiments. In: RE Spier, JB Griffiths, B Meignier (eds) 10th ESACT Meeting: Production of biologicals from animal cells in culture (pp. 609–615). Butterworth-Heinemann Ltd., Publishers, Oxford.
Mohan SB and Lyddiatt A (1991) Passive release of monoclonal antibodies from hybridoma cells. Cytotechnology 5: 201–209.
Munster MJ, Kearns MJ, Steegmans U, Behrendt U and Conner MJ (1991) A high density culture system for thein vitro production of human and mouse monoclonal antibodies. Bioprocess Eng. 6: 123–125.
Murakami H (1989) Serum-free media used for cultivation of hybridomas. In: Alan R. Liss Inc. (eds) Monoclonal antibodies: production and application (pp. 107–141).
Nakamura Y, Watanabe K, Noto T, Tajima T and Yamamura M (1989) A new compact and cell dense continuous culture system. J. Immunol. Method. 118: 31–35.
Pannell R, Milstein C (1992) An oscillating bubble chamber for laboratory scale production of monoclonal antibodies as an alternative to ascitic tumors. J. Immunol. Method. 146: 43–48.
Ronning OW, Schartum M, Winsnes A and Lindberg G (1991) Growth limitation in hybridoma cell cultures: the role of inhibitory or toxic metabolites. Cytotechnology 7: 15–24.
Schlaeger EG, Eggimann B, Gast A (1987) Proteolytic activity in serum-free culture supernatants of mouse hybridomas cells. Develop. Biol. Standard 66: 403–408.
Schlaeger EJ and Schumpp B (1989) Studies on mammalian cell growth in suspension culture. In: JB Griffiths, J Stephenne, PJ Crooy (eds) Advances in animal cell biology and technology for bioprocesses (pp. 386–396). Butterworth Publishers, Tip tree, Essex.
Schmid G, Blanch HW and Wilke CR (1990) Hybridoma growth, metabolism, and product formation in HEPES-buffered medium: I Effect of passage number. Biotechnol. Lett. 12: 627–632.
Schneider YJ (1989) Optimisation of hybridoma cell growth and antibody secretion in a chemically defined, serum-and protein-free culture medium. J. Immunol. Method. 116: 65–77.
Schumpp B and Schlaeger EJ (1989) Physiological studies of high cell density culture of different cell lines. In: JB Griffiths, J Stephenne, PJ Crooy (eds) Advances in animal cell biology and technology for bioprocesses (pp. 224–229). Butterworth Publishers. Tiptree, Essex.
Sjögren-Jansson E, Jansson G (1985) Large scale production of monoclonal antibodies in dialysis tubing. J. Immunol. Method. 84: 359–364.
Sjögren-Jansson E, Ohlin M, Borrebaeck C and Jeansson S (1991) Production of human monoclonal antibodies in dialysis tubing. Hybridoma 10: 411–419.
Sugimoto S, Lind W, Wagner R (1992) Activation of a specific proteolytic activity in suspension cultures of recombinant adherent cells. In: RE Spier, JB Griffiths, B Meignier, C Macdonald (eds) 11th ESACT Meeting: Animal cells technology: developments, process & products. (pp. 553–557). Butterworth-Heinemann Ltd., Publishers, Oxford.
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Mathiot, B., Perani, A., Dumas, D. et al. Increase of hybridoma productivity using an original dialysis culture system. Cytotechnology 11, 41–48 (1993). https://doi.org/10.1007/BF00749056
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DOI: https://doi.org/10.1007/BF00749056