Iranian Biomedical Journal

en A Simplified Process for Purification and Refolding of Recombinant Human Interferon-α2b Pharmaceutical Biotechnology Pharmaceutical Biotechnology مقاله کامل Full Length/Original Article Background: Interferon α-2b is a vital biotherapeutic produced through the recombinant DNA technology in E. coli. The recombinant IFN-α2b normally appears as intercellular IBs, which requires intensive refolding and purification steps. Method: Purification of IFN-α2b from solubilized IB was performed using two-phase extraction. To optimize refolding conditions, the effects of pH and different additives, including cysteine, cystine, urea, glycerol, Triton X-100, NaCl, and arginine, were investigated. Optimal refolding buffer (0.64 mM of urea, 5.57 mM of cysteine , and 1.8 mM of cystine) was obtained using RSM. The refolding process was performed by an optimized refolding buffer in the dilution and fed-batch refolding method at different protein concentrations (25-1000 µg/mL). Result:  At a final protein concentration of 500 µg/mL, the fed-batch refolding method yielded in a biological activity of 2.24 × 108 IU/mg, which was nearly twice that of dilution method. Conclusion: Fed-batch refolding method resulted in the biologically active IFN-α2b with high purity, which can be used for research and industrial purposes. Interferon alpha-2b, Inclusion bodies, Protein refolding 85 90 http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-4555-1&slc_lang=en&sid=1 Nima Hezarjaribi nima.hezarjaribi77@gmail.com 0000-0002-5537-5083 No Department of biology, Faculty of basic science, Islamic Azad University Science and Research Branch, Tehran, Iran Mohammad Reza Fazeli mofazeli@yahoo.com 0000-0002-6692-5028 Yes Department of Drug & Food Control, Pharmaceutical Quality Assurance Research Center, Faculty of Pharmacy, Tehran University of Medical science, Tehran, Iran