Production of aggregation prone human interferon gamma and its mutant in highly soluble and biologically active form by SUMO fusion technology》.pdf

Protein Expression and Purification xxx (2015) xxx–xxx Contents lists available at ScienceDirect Protein Expression and Purification journal homepage: /locate/yprep Production of aggregation prone human interferon gamma and its mutant in highly soluble and biologically active form by SUMO fusion technology M. Tileva a,1, E. Krachmarova a,1, I. Ivanov a, K. Maskos b, G. Nacheva a,⇑ a Institute of Molecular Biology ‘‘Roumen Tsanev”, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria b Proteros Biostructures, D-82152 Martinsried, Germany a r t i c l e i n f o a b s t r a c t Article history: The Escherichia coli expression system is a preferable choice for production of recombinant proteins. A Received 29 July 2015 disadvantage of this system is the target protein aggregation in ‘‘inclusion bodies” (IBs) that further and in revised form 16 September 2015 requires solubilisation and refolding, which is crucial for the properties and the yield of the final product. Accepted 22 September 2015 In order to prevent aggregation, SUMO fusion tag technology has been successfully applied for expression Available online xxxx of eukaryotic proteins, including human interferon gamma (hIFNc) that was reported, however, with no satisfactory biological activity. We modified this methodology for expression and purification of both the Keywords: wild type hIFNc and an extremely prone to aggregati