Abstract
A perfusion system for production of monoclonal antibodies was developed using an externally-mounted, hollow-fibre cartridge. The experimental apparatus was operated for 420 h and demonstrated increased steady-state viable cell concentration with increase in perfusion rate. Antibody titres were up to three times those measured for batch cultures and specific antibody productivity was doubled.
The procedure was successfully scaled to a 10 dm3 system which produced antibody under conditions of Good Manufacturing Practice (GMP). A calculation of productivity between the scaled perfusion system and 260 dm3 batch cultures resulted in comparable antibody production, whereas the perfusion allowed a halving in medium utilisation. Reactivity assays conducted on the purified antibody from both batch and perfusion cultures showed no evidence of proteolysis or altered antibody activity in the final perfusion product. This study provides additional support for the use of homogeneous perfusion cultures in production of monoclonal antibodies under GMP conditions.
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References
Rhodes, M.; Birch, J.: Large-scale production of proteins from mammalian cells. Biotechnology 6 (1988) 521–523
Shacter, E.: Serum-free media for bulk culture of hybridoma cells and the preparation of monoclonal antibodies. Tibtech 7 (1989) 248–253
Spier, R.: Animal cells in culture: moving into the exponential phase. Tibtech 6 (1988) 2–6
Van Brunt, J.: How big is big enough? Biotechnology 6 (1988) 480–485
Altshuler, G. L.; Dziewulski, D. M.; Sowek, J. A.; Belfort, G.: Continuous hybridoma growth and monoclonal antibody production in hollow fiber reactors-separators. Biotech Bioeng. 28 (1985) 646–658
Ku, K.; Kuo, M. J.; Delente, J.; Wildi, B. S.; Feder, J.: Development of a hollow-fibre system for large-scale culture of mammalian cells. Biotech Bioeng. 23 (1981) 79–95
Ehrlich, K. C.; Stewart, E.; Klein, D.: Artificial capillary perfusion cell culture: metabolic studies. In Vitro 14 (1978) 443–450
Klement, G.; Scheirer, W.; Katinger, H. W. D.: Construction of a large-scale membrane reactor system with different compartments for cells, medium and product. Develop Biol Stand. 66 (1987) 221–226
Shirai, Y.; Hashimoto, K.; Yamaji, H.; Tokashiki, M.: Continuous production of monoclonal antibody with immobilised hybridoma cells in an expanded bed fermentor. Appl Microbiol Biotechnol. 26 (1987) 495–499
Lydersen, B. K.; Pugh, G. G.; Paris, M. S.; Sharma, B. P.; Noll, L. A.: Ceramic matrix for large scale animal cell culture. Biotechnology 3 (1985) 63–67
Lazar, A.; Siberstein, L.; Mizrahi, A.; Reuveny, S.: An immobilised hybridoma culture system for production of monoclonal antibody. Cytotechnology 1 (1988) 331–337
Takazawa, Y.; Tokashiki, M.; Hamamoto, H.; Murakami, H.: High cell density perfusion culture of hybridoma cells recycling high molecular weight components. Cytotechnology 1 (1988) 171–178
Reuveny, S.; Velez, D.; Miller, L.; Macmillan, J. D.: Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors. J Immunol Methods, 86 (1986) 61–69
Takazawa, Y.; Tokashiki, M.; Murakami, H.; Yamada, K.; Omura, H.: High-density culture of mouse-human hybridoma in serum-free defined medium. Biotech Bioeng. 31 (1988) 168–172
Brennan, A. J.; Shevitz, J.; Macmillan, J. D.: A perfusion system for antibody production by shear-sensitive hybridoma cells in a stirred reactor. Biotech Techniques 1 (1987) 169–174
Handa-Corrigan, A.; Emery, A. N.; Speir, R. E.: Effect of gas-liquid interfaces on the growth of suspended mammalian cells: mechanisms of cell damage by bubbles. Enzyme Microb. Technol 11 (1989) 230–235
Zydney, A. L.; Colton, C. K.: A red cell deformation model for hemolysis in cross flow membrane plasmapheresis. Chem Eng Commun 30 (1984) 191–207
Lysaght, M. J.; Schmidt, M.: Factors governing mass transport in filters for membrane plasmapheresis. In: Nose, Y.; Malchesky, P. S.; Smith, J. W.; Krakauer, R. S. (Eds.): Plasmapheresis, pp. 113–127. New York: Raven Press 1983
Laugel, J.-F.; Beissinger, R. L.: Low stress shear-induced hemolysis in capillary flow. Trans Am Soc Artif Intern Organs 29 (1983) 158–162
Jaffrin, M. J.; Gupta, B. B.; Ding, L. H.; Garreau, M.: Effect of membrane dimensions and shear rate on plasma filtration for hollow fibers. Trans Am Soc Artif Intern Organs 30 (1984) 401–405
Randerson, D. H.; Blumstein, M.; Habersetzer, R.; Samtleben, W.; Schmidt, B.; Gurland, H. J.: Mass transfer in membrane plasma exchange. Artif. Organs 6 (1982) 43–49
Shiloach, J.;: Kaufman, J. B.; Kelly, R. M.: Hollow fiber microfiltration methods for recovery of rat basophilic leukemia cells (RBL-2H3) from tissue culture media. Biotech. Progress 2 (1986) 230–233
McQueen, A.; Meilhoc, E.; Bailey, J. E.: Flow effects on the viability and lysis of suspended mammalian cells. Biotech. Letters 9 (1987) 831–836
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Greenfield, P.F., Guillaume, J.M., Randerson, D.H. et al. Experience in scale-up of homogeneous perfusion culture for hybridomas. Bioprocess Engineering 6, 213–219 (1991). https://doi.org/10.1007/BF00369714
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DOI: https://doi.org/10.1007/BF00369714