Refolding, purification and characterization of the recombinant hG-CSF expressed in E. coli

Abstract

Human granulocyte colony stimulating factor (hG-CSF) is a hematopoietic growth factor produced by monocytes, fibroblasts, and endothelial cells. It stimulates the proliferation and differentiation of neutrophil precursor cells, enhancing some of the functional properties of mature neutrophils. So hG-CSF has been widely used to treat different forms of neutropenia, especially chemotherapy-induced neutropenia. In this study, we reported refolding, purification of hG-CSF expressed as inclusion body in E. coli and characterization of recombinant hG-CSF by native-PAGE and RP-HPLC chromatography showed similar yields to the standard (Neupogen). The molecular mass and peptide fragment sequence deduced from LC-MS method and the immunoassay confirmed the identity of recombinant hG-CSF. In vitro bioassay showed an equivalent biological effect (124 %) to the standard reference recombinant hG-CSF.