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Tropical Journal of Pharmaceutical Research
Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
ISSN: 1596-5996 EISSN: 1596-5996
Vol. 14, No. 3, 2015, pp. 385-390
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Bioline Code: pr15051
Full paper language: English
Document type: Research Article
Document available free of charge
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Tropical Journal of Pharmaceutical Research, Vol. 14, No. 3, 2015, pp. 385-390
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Improved Refolding Efficacy of Recombinant Human Interferon α-2b via pH Modulation
Dashbolaghi, Aziz; Khatami, Shohreh; Sardari, Sorush; Cohan, Reza Ahangari & Norouzian, Dariush
Abstract
Purpose: To increase the refolding yield of Recombinant Human Interferon α-2b in order to achieve a
highly potent product.
Methods: Interferon α-2b inclusion body was dissolved in tris-HCl buffer containing 6 M guanidine-HCl
and CuSO4. Different refolding buffers were employed for refolding the target protein. The refolded
proteins were then purified by affinity and gel filtration chromatography. The purified proteins were
subjected to circular dichroism (CD) spectropolarimetry and assayed for biological activity in vitro.
Results: Increment of pH to 8.5 improved refolding efficacies from 42.28 % to 71.22 %. However, the
relative potency significantly increased up to pH 8.0 (from 19353546 to 28633902, p < 0.05) and then
decreased to 21081305.00 at pH 8.5. The CD spectra demonstrated that by increasing pH to 8.5, the
secondary structure of the protein was altered, probably due to increase in alpha-helix from 23.7 % at
pH 7.0 to 28.1 %.
Conclusion: Employing a low-cost and simple method, such as alteration of refolding buffer pH, results
in higher refolding yield in downstream processing of rhIFN α-2b.
Keywords
Recombinant human interferon α-2b; Refolding; Circular dichroism; Spectropolarimetry; Recombinant protein; pH effect
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