Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Protein Expr Purif. 2022 Sep;197:106110. doi: 10.1016/j.pep.2022.106110. Epub 2022 May 13.
Interferon alpha-2b (IFNα-2b) is an essential cytokine widely used in hepatitis and cancer treatment. This paper presents a novel protocol for purification and efficient refolding of recombinant interferon alpha-2b (IFNα-2b) expressed as insoluble inclusion bodies in Escherichia coli. Purification of IFNα-2b from solubilized inclusion bodies was performed by solvent extraction using 2-butanol. Refolding conditions were optimized using the response surface method (RSM). Under optimized conditions, refolding yield of solvent-extracted IFNα-2b was 1.5 fold higher than refolding yield of unpurified IFNα-2b. High-concentration protein refolding was carried out by pulse-fed method, and refolding yield of 75% was achieved at a protein concentration of 300 μg ml. Under optimized conditions, 259.16 mg of purified IFNα-2b with the biological activity of 2.4 × 10 IU mg was achieved per liter of bacterial culture. The developed protocol provides an efficient production process of high-quality IFNα-2b suitable for research and pharmaceutical applications.
干扰素 alpha-2b（IFNα-2b）是一种广泛用于肝炎和癌症治疗的重要细胞因子。本文提出了一种从大肠杆菌中以不溶性包涵体形式表达的重组干扰素 alpha-2b（IFNα-2b）的新型纯化和高效复性方案。采用正丁醇溶剂萃取法从可溶包涵体中纯化 IFNα-2b。使用响应面法（RSM）优化复性条件。在优化条件下，溶剂提取 IFNα-2b 的复性产率比未纯化 IFNα-2b 的复性产率高 1.5 倍。通过脉冲进料法进行高浓度蛋白质复性，在蛋白质浓度为 300 μg/ml 时可达到 75%的复性产率。在优化条件下，每升细菌培养物可获得 259.16 毫克具有 2.4×10 IU mg 生物学活性的纯化 IFNα-2b。所开发的方案为研究和药物应用提供了一种高效的高质量 IFNα-2b 生产工艺。
