THE SIGNIFICANCE OF POLY-HIS DOMENS ON EXPRESSION LEVEL AND EFFICACY OF PURIFICATION OF HUMAN INT-A-2
Interferons (IFN) belong to cytokines family and are widely used in clinic, particularly in cancer therapy. Modern drugs fit the requirements of quality of clearing, but appear on the market with prices unacceptable for many patients. The problem of optimization of the production and purification of IFNs for simplification and cost reduction is still actual. Many laboratories are engaged in solving this problem in different countries. For production of highly purified substances, we used pET bacterial expression system (Escherichia coli) for production of a human IFN-a-2b. The genetic design containing synthesized and modified sequences of target protein has been cloned. We have synthesized three genetic designs of hIFN-a-2b: without histidine tag and with domains consist of 6 or 12 histidine (6-His and 12-His). Proteins expressed were identified as IFN-a-2b using ELISA and Western blotting assaus. Our data indicate that the design with the 6His on the N-end is the most successful construction either the level of expression or efficacy of purification.