Recombinant human G-CSF production as a protein based drug candidate for hematology and oncology

Abstract

The human granulocyte colony stimulating factor (hG-CSF) is a member of the CSF family. The first purpose of this study was production of recombinant human G-CSF (rhG-CSF) which is an important therapeutic protein for angiogenesis based clinical applicaitons. rhG-CSF was produced as inclusion body in the Escherichia coli strain BL21 (DE3) pLysE with pTOLT vector system. The rhG-CSF was purified by Ni-NTA agarose afinity chromatography and characterized with SDS-PAGE analysis. The effects of this therapeutic protein on cell viability was measured by MTT assay. Additionally, angiogenic potential of produced rhG-CSF was investigated via HUVEC cell line by in vitro scratch assay. As a result, the purified protein induced cell proliferation and the EC50 value of protein based drug candidate was 0.051 mM. Additionally, it was determined that the migration ability of the HUVECs was promoted by rhG-CSF in a concentration-dependent manner by in vitro scratch assay.