Abstract
The combination of BD Cell MAb medium with the CL-1000 flask is increasingly being used to generate a few hundred milligram of antibody for early stage research projects. Cells are inoculated at 2 million per ml, and the antibody is harvested after 15 days or when the antibody concentration reaches above 10 mg ml−1, whichever comes first. Currently, there is no means to scale up beyond this production level using this technology. In this study, we evaluated hollow fiber technology as the scale up alternative. The hollow fiber system was run in batch mode to mimic the method used for the CL-1000 with BD MAb medium. The FL-NS murine hybridoma cell line was simultaneously inoculated at 2 million cells per ml in a CL-1000 and the Maximizer hollow fiber bioreactor system, a 21-fold theoretical scale up over the CL-1000. The Maximizer produced 23-fold more antibody, very close to the expected theoretical amount. However, production was complete after 9 days in the Maximizer, while the CL-1000 required the full 15 days for production. In summary, these results demonstrate successful scale up of antibody production from the CL-1000 to a hollow fiber system.
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References
Fritchman K. 2000. New cell culture medium scores high marks for yield and versatility. Pharmaceutical Manufacturing International, 29 pp.
Gramer M.J. and Britton T.L. 2002. Antibody production by a hybridoma at high cell density is limited by two independent mechanisms. Biotechnol. Bioeng. 79: 277–283.
Gramer M.J., Poeschl D.M., Conroy M.J. and Hammer B.E. 1999. Effect of harvesting protocol on performance of a hollow fiber bioreactor. Biotechnol. Bioeng. 65: 334–340.
Gramer M.J. and Poeschl D.M. 1998. Screening tool for hollow fiber bioreactor process development. Biotechnol. Prog. 14: 203–209.
Hirschel M.D. and Gruenberg M.L. 1988. An automated hollow fiber system for the large scale manufacture of mammalian cell secreted product. In: Lydersen B.K. (ed.), Large Scale Cell Culture Technology, Macmillan, pp. 113–144.
Miller W.M., Blanch H.W. and Wilke C.R. 1988. A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: effect of nutrient concentration, dilution rate, and pH. Biotechnol. Bioeng. 32: 947–965. PhRMA Survey, 2002. New Medicines in Development for Biotechnology www.phrma.org.newmedicines/resources/ 2002-10-21.93.pdf.
Piret J.M. and Cooney C.L. 1991. Model of oxygen transport limitations in hollow fiber bioreactors. Biotechnol. Bioeng. 37: 80–92.
Trebak M., Chong J.M., Herlyn D. and Speicher D.W. 1999. Efficientlaboratory-scaleproductionofmonoclonalantibodies using membrane-based high-density cell culture technology. J. Immunol. Meth. 230: 59–70.
Wolf M.L. and DeSutter T. 1999. High density cell culture in a new passive membrane based bioreactor. In: Conner T.H., Weier H.-U. and Fox F. (eds), Biotechnology International II, Universal Medical Press, San Francisco, CA, pp. 293–301.
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Gramer, M.J., Maas, J. & Lieberman, M.M. Use of hollow fiber systems for rapid and direct scale up of antibody production from hybridoma cell lines cultured in CL-1000 flasks using BD Cell MAb medium. Cytotechnology 42, 155–162 (2003). https://doi.org/10.1023/B:CYTO.0000015842.54956.24
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DOI: https://doi.org/10.1023/B:CYTO.0000015842.54956.24