A method of purification of the recombinant protein products.

Abstract

The present invention relates to development of a methods of purification of the recombinant protein products which comprises of the following steps: a. culturing recombinant strains of E.coli or Pichia pastor is in respective growth media, b. fermentation with agitation of 250-600 rpm was allowed to proceed for 20 hours for E.coli or 105 hours for Pichia pastoris, c. harvesting by centrifugation at 4°C and 4200 g for 20 minutes, d. re-suspending the pellet at 4°C in Tris-HCI buffer pH8, using 8.0ml of buffer for each gram of cells, disrupted by passing the re-suspended cell solution two times through the French-press at a maximum pressure of 1.6 K.bar at 4°C. Inclusion bodies (IB) was separated by centrifuging at 16.200g for 30 min at 4°C. Pellet was resuspended in Tris-HCI buffer and recovered by centrifugation; e. re-suspending the pellet in the Solublization buffer (SB) (Guanidine - Tris HCI, pH 8) until the solution became clear, centrifuged at 16.200g for 30 minutes at f. 4°C. Interferon protein was recovered at 4.7mg / ml; g. diluting the protein solution into a larger volume of the refolding solution containing O.SM L-arginine, 100mM Tric-HCI and 2mM EDT A pH8, in the presence of both reducing loxidizing agents; h. dissolving inclusion bodies was gently poured ill to the refolding buffer and agitated for 30 hours under constant stirring and temperature.