For over ten years, recombinant human Granulocyte Colony Stimulating Factor (hG-CSF) has been
broadly used in clinical applications. Many sources of G-CSF have the disadvantage of being in
intracellular or aggregated form. In this paper, we report the molecular cloning and gene expression in the
protease resistant strain of yeast (SMD1168). More than 90% of the G-CSF protein is in secreted form and
about 100 mg of the biologic active human G-CSF can be obtained from two liters of bioreactor. The recombinant G-CSF promotes HL-60 (Human promyelocytic cell) cell proliferation and elevates the mito-chondrial succinate dehydrogenase activity. The dose effects of rG-CSF on cell proliferation responses to the total numbers of the receptors on the cell surfaces
have been investigated and the quantitative results were obtained. The receptors are saturated byG-CSF atthe50 ng/mLfor 1x
10^5 cells. The prompt increasing in the succinate dehydrogenas activity was also observed. This effect is comparable to that from the commercial source. We have successfully cloned and expressed a bioactive rhG-CSF from the medium in
the Pichia pastoris system. This specific product is expected to meet the clinical applications in the treatments of neutropenia, acute cerebral ischemia and other neuro-degenerated diseases.
關聯:
Journal of the Chinese Chemical Society=中國化學會會誌 57(4B), pp.850-856
