Enhancing recombinant interleukin‐6 production yield by fermentation optimization, two‐step denaturing, and one‐step purification. (23rd September 2017)

• Record Type: Journal Article
• Authors: Ahmed, Nadeem; Abbas, Rabbia; Khan, Mohsin Ahmad; Bashir, Hamid; Tahir, Saad; Zafar, Ahmad Usman
• Is Part Of: Biotechnology and applied biochemistry. Volume 65:Number 3(2018)
• Page Start: 490
• Page End: 496
• Publication Date: 2017-09-23
• Subjects: two‐step denaturing -- one‐step purification -- rhIL‐6; Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1588
• Languages: English
• ISSNs: 0885-4513

Abstract

Abstract: Interleukin‐6 a pleiotropic cytokine involved in a wide range of biological activities. So the large‐scale production of biologically active recombinant human interleukin‐6 is important for its structural and functional studies. Here, we report an optimized method for shake flask fermentation and a simplified high‐yield purification procedure for the recombinant interleukin‐6. This high‐yield expression method not only involves the optimization of the fermentation condition but also the single step purification method as well as a two‐step denaturing and one‐step refolding process. This approach replaces the more conventional procedure of protein solubilization and refolding. Through applying these strategies, the final cell density and overall product yield of the recombinant human interleukin‐6 were obtained as 20.4 g as cell biomass and 150 mg as purified active protein from the I‐L of the culture. The purified protein was characterized by HPLC and SDS–PAGE. The results of the current work demonstrate that the described method may be used to develop the process for industrial‐scale production of the biologically active recombinant interleukin‐6 protein.