STUDY ON INFLUENCE OF ADDITIVES ON REFOLDING AND DIALYSIS OF RECOMBINANT HUMAN INTERFERON GAMMA
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Keywords
rhIFN-γ, protein expression, refolding, dialysis, additives.
Abstract
Recombinant protein accumulated intracellularly is frequently laid down in the form of inclusion bodies. So, dialysis and refolding condition optimization can be advantageous to standardize purification protocol with minimum protein yield loss. Recombinant Human interferon gamma (rhIFN-γ) IB was solubilized at alkaline pH (8.0) in 6 M Guanidinium HCl solution. The solubilized proteins were checked for protein loss when refolded and dialyzed at different buffer pH and in presence of different additives like ßME (â-Mercapto Ethanol), Tween-20, Mannitol and their combination. Phosphate buffer at 50mM concentration (pH 6.5) minimized the protein loss. Tween- 20 (0.01% W/V) increased recovery of protein from inclusion bodies.
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D.M. Shukla
Gujarat Pollution Control Board, Gandhinagar, Gujarat 382010, India.
