Demand for increasingly complex post-translationally modified proteins, such as monoclonal antibodies (mAbs), necessitates the use of mammalian hosts for production. The focus of this article is a continuous centrifugal bioreactor (CCBR) capable of increasing volumetric productivity for mAb production through high density hybridoma culture, exceeding 10(8) cells/mL. At these extreme densities, environmental conditions such as substrate and inhibitor concentrations rapidly change dramatically affecting the growth rate. The development of a kinetic model predicting glucose, mAb, lactate, and ammonium concentrations based on dilution rate and cell density is shown in this article. Additionally, it is found that pH affects both growth rate and viability, and a range of 6.9-7.4 is needed to maintain growth rate above 90% of the maximum. Modeling shows that operating an 11.4 mL CCBR inoculated with 2.0 x 10(7) cells/mL at a dilution rate of 1.3 h(-1), results in a predicted growth rate 82% of the maximum value. At the same dilution rate increasing density to 6.0 x 10(7) cells/mL decreases the predicted growth rate to 60% of the maximum; however, by increasing dilution rate to 6.1 h(-1) the growth rate can be increased to 86% of the maximum. Using the kinetic model developed in this research, the concentration of glucose, mAb, lactate, and ammonium are all predicted within 13% of experimental results. This model and an understanding of how RPM impacts cell retention serve as valuable tools for maintaining high density CCBR cultures, ensuring maximum growth associated mAb production rates.
(c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.