Chromatographic Purification and Refolding of an Insoluble Histidine Tag Recombinant Human Interleukin-2 Expressed in E. coli

Recombinant human interleukin-2 was expressed as inclusion body in E.coli. An ultrasonic homogenization was done for cell disruption and then inclusion bodies isolated by centrifugation. The inactive inclusion bodies were solubilized in 8 M urea / 20 mM β-mercaptoethanol. Purification and refolding was done on immobilized metal-ion affinity chromatography (IMAC) in one step. A gradual refolding of the protein was performed on-column from 8 M to 0 M urea. The refolded protein was then eluted by refolding buffer containing 0.5 M urea and 300 mM imidazole. Then activity of the purified protein was investigated. It indicated that purified IL-2 can proliferate the T cells.

Keywords:

Recombinant Human Interleukin-2 , Inclusion Body , IMAC , Refolding