Abstract
Proteolytic enzymes existing in plant cell cultured media are the major reason for the loss of secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The addition of pepstatin, aprotinin and PMSF relatively decreased the proteolytic degradation of hGM-CSF in a conditioned medium, but sufficient prevention against the proteolytic activity could not be obtained with chemical protease inhibitors. Gelatin, as a competitive substrate for protease, showed a stabilizing effect in a conditioned medium. Compared to the initial hGM-CSF concentration in a conditioned medium, with 10 g/L of gelatin, 68% of the hGM-CSF remained after 5 days. In a cell culture experiment, 5 g/L of gelatin significantly stimulated the hGM-CSF production and accumulation in culture media, with no growth inhibition. Compared to the controls (4.72 μg/L), the extracellular hGM-CSF level could be increased to 39.78 μg/L with the addition of 5 g/L of gelatin.
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Lee, SY., Cho, JM. & Kim, DI. Stability enhancement of hGM-CSF in transgenicNicotiana tabacum suspension cell cultures. Biotechnol. Bioprocess Eng. 8, 187–191 (2003). https://doi.org/10.1007/BF02935895
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DOI: https://doi.org/10.1007/BF02935895