Abstract
The performance of a hollow-fiber membrane bioreactor (HFBR) (molecular weight cut-off 30 kD, fiber surface area 2050 cm2) containing a culture of hybridoma cells has been investigated. Experimental data were used as basis to develop a model of general application. Concentrations of fundamental nutrients (glucose and glutamine), inhibitory products (ammonium and lactate), and monoclonal antibodies (MAb) against bovine lactoferrin (IgG1) were monitored over time. Exchange of nutrients and products occurred across the capillary surface, whereas cells and MAb remained in the extra-capillary space (ECS). A protein-free culture medium (Hybrimax) with and without antibiotics was used. In both cases, the final MAb concentration was the same; however, antibiotic presence slowed down the time to achieve this concentration. Diffusion assays have been carried out in order to support the development of a mathematical model that describes the performance of the HFBR, including mass transfer and reaction terms. Inhibition by ammonium and lactate has been considered in the kinetics, providing model results consistent with experimental data. Further research with other cell lines and/or culture media will allow to broaden the field of application of this model for general use in HFBR systems.
Highlights
Hybridoma technology was developed to overcome the limitations of polyclonal antibodies[1]
This paper has developed an applicable reaction-diffusion model, assuming certain simplifications based on additional experiments, to assess the performance of the system, i.e. conversion of substrates into products, as well as Monoclonal antibodies (MAb) production with and without antibiotics
Glucose and glutamine concentrations decreased throughout the duration of the experiment, the consumption rate was slower during the last hours of experimentation
Summary
Hybridoma technology was developed to overcome the limitations of polyclonal antibodies (broad specificity, possible cross-reactivity or variability among immune serum batches)[1]. The growing demand for monoclonal antibodies at reasonable prices means that it is necessary to carry out an optimization of their production processes[2]. Since the development of cell hybridoma technology, production of ascites in mice has been the primary method available for the production of large amounts of monoclonal antibodies[3]. Analysis of the evolution of substrate and product concentrations is fundamental in the determination of hybridoma kinetics. Glucose and glutamine are key substrates for hybridoma growth. Part of the glutamine is deaminated, yielding ammonium and glutamate, which is later transformed into other amino acids for biosynthesis purposes. Whereas glucose and glutamine are fundamental nutrients, ammonium and lactate are products of cell metabolism that can act as inhibitors when their concentrations are high enough[6]. Due to increasing safety concerns as well as cost issues, the requirements for biotechnological
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