Enhanced production of recombinant proteins from plant cells by the application of osmotic stress and protein stabilization.
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.
Soderquist
Ryan G
RG
Department of Chemical Engineering, Washington State University, Pullman, WA 99164-2710, USA. rsoder@wsu.edu
Lee
James M
JM
D000383:Agriculture / Q000379:methods; D002478:Cells, Cultured; D045604:Extracellular Fluid / Q000187:drug effects / Q000502:physiology; D005780:Gelatin / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D008353:Mannitol / Q000494:pharmacology; D009997:Osmotic Pressure / Q000187:drug effects; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology; D014882:Water-Electrolyte Balance / Q000187:drug effects / Q000502:physiology
D011993:Recombinant Fusion Proteins; D012710:Serum Albumin, Bovine; D008353:Mannitol; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D005780:Gelatin