Optimization of culture conditions for high-level expression of soluble and active tumor necrosis factor-? in E. coli

Anti-TNF inhibitors exert their therapeutic effect by inhibition of the excessive amounts TNF-? within body. Recombinant should be produced in a soluble refolded form to investigate effectiveness and efficiency anti-TNF-? compounds. In this research, designed cassette was subcloned pET28a expression vector expressed E. coli BL21 (DE3). The identity protein confirmed through SDS-PAGE Western blotting. After optimizing conditions, purification performed using native Ni-NTA affinity chromatography. biological activity recombinant investigated MTT assay. Also, an agent, Altebrel, against ELISA. Optimization conditions represented that highest could achieved at 37 °C 0.5 mM IPTG 6 h post-induction. inhibitory on L929 murine fibroblast cell line successfully detected Altebrel Binding kinetics were also studied Cimzia as molecule 7.2 E?13M calculated equilibrium dissociation constant value (KD). significant level form, its high purity, assessment showed used tests ELISA assess agents.