Kinetic characterization of metabolism and antibody production of a hybridoma cell line in high density homogeneous perfusion culture

Since the advent of monoclonal antibodies (MAbs) from hybridoma cells in 1975, hybridoma technology has had a major impact on the scientific and research world. The demand for MAbs has increased considerably in recent years. MAbs are being used in in vitro diagnostics, affinity purification, in vivo imaging and immunotherapy. While the demand for MAbs will remain primarily stable in other areas, it is projected that human therapy (primarily the treatment of cancer) will require large quantities of highly pure monoclonal antibodies, which can be conservatively estimated at 50 kilograms/year for each therapeutic MAb. The excessive costs of producing sufficient quantities of MAbs and the small yields from conventional technology are important factors in making research and development of cost effective large scale MAb production essential. From a biochemical engineering perspective, to increase any bioreactor performance the product of the viable cell density (cells/ml) and the specific MAb production rate (production rate per cell) has to be maximized.;The goal of this research was to compare cell growth, antibody production and metabolic rates of hybridoma cells in suspension culture over a range of cell densities and specific growth rates; and, to investigate the effects of these variables on the environmental parameters such as nutrient and product concentrations. Since there are few reports on the metabolism of hybridoma cells at high densities, much of the effort has been aimed at gathering information on cell behavior at such high densities and to do an analysis of metabolic, growth and production rate data of high density perfusion systems.;A homogeneous perfusion reactor was developed to operate continuously at suspended cell densities greater than ;The results show that the specific MAb production rate is a stronger function of cell density than of specific growth rate with the specific MAb production rate increasing with viable cell density. There are also other favorable shifts in the energy metabolism as the cell densities increase. The developed reactor has been found to be very effective for long term culture of hybridoma cells when operated in a partial cell retention mode. This study increases our knowledge of the energetics and regulation of cell growth, metabolism and production rates in high density, homogeneous perfusion reactors.