| 摘要 |
PURPOSE:To carry out the purification of interferon in high efficiency, suppressing the denaturation of polypeptide and the decomposition with protease, by adding protamine to the cultured product of microorganism capable of producing physiologically active polypeptide and obtained by a recombinant DNA technique, and carrying out the extraction and purification of the objective interferon. CONSTITUTION:A microbial strain transformed by a plasmid vector containing a gene coding a physiologically active polypeptide [e.g. human immunity interferon (h-INF-gamma)-producing Escherichia coli W3110/PIN5T4N143, etc.] is cultured in a medium containing e.g. 3(wt)% polypeptone, 2% yeast extract, 2% glucose, 0.5% KH2PO4, 0.01% MgSO4.7H2O, and 20mug/ml of tetracycline, at 30 deg.C for 24hr under aeration and agitation, and the proliferated cells are separated by centrifugal separation. The cells are suspended in a tris-hydrochloric acid buffer solution of 7.5pH, added with protamine, disintegrated under ice-cooling, and centrifuged to obtain supernatant liquid. The supernatant liquid is purified e.g. by ammonium sulfate fractionation, chelate column fractionation, etc. |
