title: Optimized conditions for high-level expression and purification of recombinant human interleukin-2 in E. coli.
abstract: Interleukin-2 (IL-2), a potent cytokine has been used in anti-cancer therapy for over a decade now. IL-2, originally identified as a growth factor for T lymphocytes is a 15 kDa hydrophobic glycoprotein that induces the activation, clonal proliferation and differentiation of T and B-lymphocytes and enhances the cytotoxicity of monocytes and natural killer (NK) cells. Here, we report a simple method for the cloning, high-level expression and purification of IL-2 protein, which can be easily extended to other bioactive therapeutic proteins. The IL-2 gene was amplified from human spleen cDNA and cloned in a prokaryotic (E. coli) expression system. An optimal expression of the IL-2 protein was determined by varying the expression conditions like temperature, inducer concentration and duration of induction. The protein was expressed as inclusion bodies and a panel of reagents including detergents, urea and guanidine hydrochloride were used to solubilize it. After solubilization, the protein was renatured and subjected to a single step gel-filtration chromatography to yield immunobioactive IL-2 protein with > 99% purity.
authors.affiliation: Dabur Research Foundation, 22, Site IV, Sahibabad, Ghaziabad, Uttar Pradesh 201010, India.
authors.forename: Paromita
authors.initials: P
authors.lastname: Sengupta
authors.forename: Kalpana
authors.initials: K
authors.lastname: Meena
authors.forename: Rama
authors.initials: R
authors.lastname: Mukherjee
authors.forename: S K
authors.initials: SK
authors.lastname: Jain
authors.forename: Kapil
authors.initials: K
authors.lastname: Maithal
mesh_terms: D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D007376:Interleukin-2; D011994:Recombinant Proteins