A Simplified Process for Purification and Refolding of Recombinant Human Interferon_a2b

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Hezarjaribi Nima; FAZELI MOHAMMAD REZA

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

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Journal: Iranian Biomedical Journal Year:2022 | Volume:26 | Issue:1 Page(s): 85-90

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Title

A Simplified Process for Purification and Refolding of Recombinant Human Interferon_a2b

Author(s)

Hezarjaribi Nima | FAZELI MOHAMMAD REZA | Issue Writer Certificate

Keywords

Interferon alpha-2b

Inclusion bodies

Protein refolding

Abstract

Background: Interferon α-2b is a vital biotherapeutic produced through the recombinant DNA technology in E. coli. The recombinant IFN-α 2b normally appears as intercellular IBs, which requires intensive refolding and purification steps. Method: Purification of IFN-α 2b 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 (251000 µ g/mL). Result: At a final protein concentration of 500 µ g/mL, the fed-batch refolding method yielded in a biological activity of 2. 24 × 10 8 IU/mg, which was nearly twice that of dilution method. Conclusion: Fed-batch refolding method resulted in the biologically active IFN-α 2b with high purity, which can be used for research and industrial purposes.

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APA: Copy

Hezarjaribi, Nima, & FAZELI, MOHAMMAD REZA. (2022). A Simplified Process for Purification and Refolding of Recombinant Human Interferon_a2b. IRANIAN BIOMEDICAL JOURNAL, 26(1), 85-90. SID. https://sid.ir/paper/988107/en

Vancouver: Copy

Hezarjaribi Nima, FAZELI MOHAMMAD REZA. A Simplified Process for Purification and Refolding of Recombinant Human Interferon_a2b. IRANIAN BIOMEDICAL JOURNAL[Internet]. 2022;26(1):85-90. Available from: https://sid.ir/paper/988107/en

IEEE: Copy

Nima Hezarjaribi, and MOHAMMAD REZA FAZELI, “A Simplified Process for Purification and Refolding of Recombinant Human Interferon_a2b,” IRANIAN BIOMEDICAL JOURNAL, vol. 26, no. 1, pp. 85–90, 2022, [Online]. Available: https://sid.ir/paper/988107/en

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