Enhanced production of recombinant proteins from plant cells by the application of osmotic stress and protein stabilization

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Human granulocyte-macrophage colony-stimulating factor (GM-CSF) was produced from transgenic Nicotiana tabacum cells. The application of osmotic stress through the addition of 90 g/l mannitol to the plant cell medium enhanced the maximum extracellular GM-CSF concentration from 76 microg/l to 130 microg/l (1.7-fold increase). The addition of bovine serum albumin (BSA), along with mannitol, further increased the maximum extracellular GM-CSF concentration by as much as 2.5-fold over the control. GM-CSF degradation studies in conditioned medium demonstrated that mannitol and BSA both stabilize the GM-CSF protein. The addition of gelatin together with mannitol to the plant cell medium also enhanced the maximum extracellular GM-CSF concentration and stability over time.