Production of Functional Recombinant Human Interferon-gamma by RTX CPD-fusion Technology

Human interferon-gamma (hIFNγ) is an important pleiotropic cytokine with therapeutic application. Recombinant hIFNγ has been shown to be effective pharmaceutical against a wide range of viral, immuno-suppressive diseases promising prospects in cancer immunotherapy resulting strong increase demand and price. The aim this study was develop efficient method for production soluble active hIFNγ. To end fusion gene encoding C-terminally located His10-tagged RTX CPD constructed by two-step PCR. cloned expression vector under inducible promoter expressed different E. coli strains such as Rosetta (appropriate eukaryotic genes containing rare codons), two chaperone – BL21(DE3)/pG-KJE8 BL21(DE3)/pG-Tf2. transformed bacteria were cultivated at 24℃ favour proper folding the recombinant protein. Under these conditions strain chosen more suitable latter purified Ni2+- based affinity chromatography, peak fractions subjected chromatography combined direct on-column digestion release hIFNγ, which further ion exchange chromatography. It worth mentioning that strategy, employing auto-cleaving solubility-enhancing tag, tag applied first time