Summary

Research and Development of Therapeutic Recombinant Human Granulocyte Colony Stimulating Factor

hG-CSF (human Granulocyte Colony-Stimulating Factor), consisting of 174 amino acid, is a colony-stimulating factor hormone, to stimulate the bone marrow to produce granulocytes and stem cells. G-CSF then stimulates the bone marrow to release them into the blood. hG-CSF was approved to treat for cancer patients receiving myelosuppressive chemotherapy, patients with acute myeloid leukemia receiving induction or consolidation chemotherapy patients with severe chronic neutropenia. In this study, gene endoing for hG-CSF was cloned to pNanogenGCSF1 expression vector and transformed to E.coli to create E.coliBL21(DE3) GCSF1 strain used to overexpress hG- CSF in LB medium with 0.4%glucose, 0.5mM IPTG, 250rpm, 37⁰C, 6 hours. Extraction and purification of hG-CSF procedure archived the recombinant hG-CSF with purity of >95%, total protein concentration >1mg/ml, pH 3.5-4.5. The purified protein has molecular weight of 18-20 KDa, no host cell derived DNA and peptide residue contamination; it is also endotoxin free and has the biological activity of 101.13 MU/mg.

Keywords: G-CSF, cloning, expression, E.coli, neutropenia.