Thesis
Enhanced production of recombinant proteins from plant cells by the application of osmotic stress
Master of Science (MS), Washington State University
2004
Handle:
https://hdl.handle.net/2376/169
Abstract
Transgenic plant cell suspensions offer several unique advantages for the production of recombinant proteins. This work has shown that osmotic stress and protein stabilization can significantly enhance recombinant protein production from a plant cell suspension. Transgenic Nicotiana tabacum cells producing human granulocytemacrophage colony-stimulating factor (GM-CSF) were used as the model system for this work. The application of osmotic stress with mannitol at a concentration of 90 g/L enhanced the maximum attainable GM-CSF concentration by 1.7-fold. Mannitol addition also enhanced the ratio of total protein secretion per gram of cells by 3.5-fold. The addition of bovine serum albumin (BSA) along with mannitol in a time-course manner increased the maximum attainable GM-CSF concentration by 2.5-fold over the control. BSA and mannitol addition also stabilized GM-CSF at concentrations of over 100 ng/ml for several days. Studies on GM-CSF degradation in the media revealed that both mannitol and BSA stabilized GM-CSF independently. Degradation studies also demonstrated that there are different stabilization effects between commercially supplied and plant-produced GM-CSF. Sodium chloride and BSA were not able to interact as effectively as mannitol and BSA to improve the GM-CSF yield. The unique ability of mannitol to interact with a secondary stabilizer is an important finding that can enhance recombinant protein yields.
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Details
- Title
- Enhanced production of recombinant proteins from plant cells by the application of osmotic stress
- Creators
- Ryan G. Soderquist
- Contributors
- James M. Lee (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
- Identifiers
- 99900525061101842
- Language
- English
- Resource Type
- Thesis