Thesis
Enhanced production considering temperature effects on new hybridomas for bovine natural killer cell monoclonal antibodies
Washington State University
Master of Science (MS), Washington State University
2008
Handle:
https://hdl.handle.net/2376/101467
Abstract
Demand for monoclonal antibodies (mAbs) has led to the improvement of mammalian cell culture processes taking into account reactor design and specific mAb production rate of the cell line. The creation of a continuous centrifugal bioreactor (CCBR) allows for increased volumetric productivity by maintaining a high density cell culture, 108 cells/mL, while lowering the cell culture temperature increases the specific mAb production rate. This thesis contains a manuscript prepared for submission to Journal of Veterinary Immunology and Immunopathology in which I describe the creation of new hybridoma cell lines secreting mAb to bovine natural killer (NK) cells. The recent development of a mAb to the bovine orthologue of the activating NK receptor NKp46 allowed us to develop a flow cytometric method for clustering and characterizing additional mAbs reactive with bovine NK cells. Cluster 1 mAbs, NK64A and NK93A, recognize an orthologue of the stress protein gp96 expressed on NK cells and granulocytes. Cluster 2 mAbs, NK29A, NK42A, NK47A, NK86A, NK134A, and NK137A, recognize a yet undetermined molecule expressed on activated NK cells and subsets of [alpha][Beta] and [gamma][delta] T cells. The NK64A hybridoma was utilized in studies of the effect of cell culture temperature on cell growth and specific mAb production using the CCBR and a kinetic model for predicting glucose, ammonium, lactate, and mAb concentrations. Batch studies were performed at temperatures of 31, 33, 35, 37, and 39°C; and for the temperature range 35 to 39°C it was found that the growth rate was essentially constant, 0.052 hr-1 ± 0.001, but decreased to 0.036 hr-1 for 33°C and 0.026 hr-1 for 31°C. The specific mAb production rate increased by 120% at 33°C and 100% at 31°C compared to 37°C. Culture of the NK64A hybridoma in the CCBR at 37 and 33°C resulted in decreased growth rates compared to batch studies, 0.0039 hr-1 and 0.011 hr-1 respectively, indicating cell density inhibition for this cell line. Yield coefficients were adjusted to improve fit of the kinetic model to the CCBR data. These results indicate the need to study each cell line in order to determine the optimum cell culture conditions.
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Details
- Title
- Enhanced production considering temperature effects on new hybridomas for bovine natural killer cell monoclonal antibodies
- Creators
- Aella Natasha Godwin
- Contributors
- Bernard J. Van Wie (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Chemical Engineering and Bioengineering, School of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; [Pullman, Washington] :
- Identifiers
- 99900525294401842
- Language
- English
- Resource Type
- Thesis