Soluble Expression of IFNα2-Tα1 Fusion Protein in Escherichia coli by N-terminal SUMO Fusion and its Anti-Proliferative Activity
Soluble Expression of IFNα2-Tα1 Fusion Protein in Escherichia coli by N-terminal SUMO Fusion and its Anti-Proliferative Activity
Muhammad Shahbaz Aslam1, Iram Gull1, Zaigham Abbas2,* and Muhammad Amin Athar1
ABSTRACT
Interferon alpha 2 (IFNα-2) is a type of cytokine with both antiviral and anticancer activities. Binding of IFNα-2 to its receptor on cell surface leads to activation of interferon stimulated genes which mediate its anti-proliferative and anti-angiogenic properties. Similarly, Thymosin alpha 1 (Tα-1) helps to fight against different infections such as cancer and hepatitis with its immune modulating properties as well as through its direct action on target cells. The recombinant expression of some proteins in E. coli produces inclusion bodies which are misfolded proteins. The objective of this study was soluble expression of IFNα2-Tα1fusion protein in E. coli using pET-SUMO vector and determination of its biological activity. SUMO-IFNα2-Tα1 was successfully expressed in soluble form with IPTG induction to final concentration 0.5 mM at 37oC for 4 h and purified using affinity chromatography. SUMO tag was removed from SUMO-IFNα2-Tα1 by SUMO protease and recombinant IFNα2-Tα1 was collected in flow through by affinity chromatography. The MW (~23 kDa) of IFNα2-Tα1 was determined by 12 % SDS-PAGE and its integrity was confirmed by Immuno blot analysis using anti-interferon α-2 and anti-thymosin α-1 antibodies. Purified IFNα2-Tα1 demonstrated anti-proliferation activity as assessed by MTT assay. This study also showed that N-terminal fusion of SUMO withIFNα2-Tα1 is effective for its soluble expression and to make its purification process more convenient.
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August 2018
Vol. 50, Iss. 4, Pages 1199-1600
