Recovery of active anti TNF-alfa ScFv through matrix-assisted refolding of bacterial inclusion bodies using CIM monolithic support

K. Sushma, C. J. Bilgimol, M. A. Vijayalakshmi, P. K. Satheeshkumar

Journal of Chromatography B, 891 - 892 (2012) 90 - 93(2012) 90 - 93

Anti TNF-α molecules are important as therapeutic agents for many of the autoimmune diseases in chronic stage. Here we report the expression and purification of a recombinant single chain variable fragment (ScFv) specific to TNF-α from inclusion bodies. In contrast to the conventional on column refolding using the soft gel supports, an efficient methodology using monolithic matrix has been employed. Nickel (II) coupled to convective interaction media (CIM) support was utilized for this purpose with 6 M guanidine hydrochloride (GuHCl) as the chaotropic agent. The protein purified after solubilization and refolding proved to be biologically active with an IC50.

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