Isolation of human interferon β1b (Ser17) expressed in E. coli with the use of ion-exchange chromatography

• Published in: Russian journal of bioorganic chemistry 2011-05, Vol.37 (3), p.292-297
• Main Authors: Romanov, V. P., Bezuglov, V. V., Bobrov, M. Yu, Kostromina, T. I., Feofanov, S. A., Miroshnikov, A. I.
• Format: Article
• Language: English
• Subjects: Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Life Sciences; Organic Chemistry
• ISSN: 1068-1620; 1608-330X
• DOI: 10.1134/S1068162011030150

A method for isolation of interferon β1b (Ser17) from the inclusion bodies is presented. This method consists of the following stages: solution and reduction of the protein from the inclusion bodies, refolding, chromatography on DEAE-sepharose, chromatography on SP-sepharose, concentration of the protein solution, demineralization, and addition of stabilizers. The refolding was performed by dilution of the solution of the reduced protein with a buffer (pH 8.0) containing 50 mM Tris-HCl, 25 μM CuCl 2 , and 0.5% Tween-20. Interferon β1b was eluted from a cation-exchange column with a pH gradient (9.3–11.3) of sodium phosphate buffer. The eluate was concentrated and desalted on a column with sephadex G-50 in 1 mM NaOH, mannitol, and sodium phosphate were added for neutralization of the protein solution. The product was homogenous according to gel electrophoresis and HPLC, and exhibited the practically same antiproliferative activity as that of Betaferon taken as a standard. Thus, a possibility of preparation of the purified and active interferon β by ion-exchange chromatography in the presence of the Tween-20 nonionic detergent was demonstrated. The proposed procedure is promising for application to large-scale production and industry.