FIELD: biotechnology; genetic engineering.
SUBSTANCE: invention relates to a medicinal product, which is a solution for parenteral administration containing osmotic pressure regulators, a buffer system maintaining a pH of 4.0–6.5, Tween 20 and highly purified interferon gamma (INF -gamma) with amino acid sequence SEQ ID NO: 1, wherein interferon gamma is obtained by cultivating the producer strain of Escherichia coli JM109/pET32a-IFG144, where after 3–5 hours of cultivation the biosynthesis of the recombinant protein is induced in Escherichia coli cells JM109/pET32a-IFG144 isopropyl-β -D-1-thiogalactopyranoside and incubated for 4–6 hours, obtaining cell biomass of the producer strain producing the INF-gamma protein, isolating the IFN-gamma protein-producing strain from the resulting cell biomass, subsequent purification of the isolated IFN-gamma protein by cation exchange chromatography under denaturing conditions, renaturation of the IFN-gamma protein by sequential fractional addition of the denatured protein to a renaturation buffer containing L-arginine, and purification of the renatured IFN-gamma protein by cation exchange chromatography, where interferon gamma has a purity of at least 97% and a specific biological activity of at least 1.7×107 IU per 1 mg of protein, and the producer strain was obtained by transforming the parent strain E. coli JM109 with the recombinant plasmid pET32a-IFG144 of 5806 base pairs in size, containing the structural elements: T7 promoter and lacO operator, synthetic nucleotide sequence SEQ ID NO: 2, encoding the INF-gamma protein, the ampicillin antibiotic resistance gene (AmpR) and the bacterial promoter of the ampicillin resistance gene (AmpR promoter) for selection of recombinant cells, the origin of replication of bacteriophage f1 (f1 ori), the lactose repressor gene LacI and the bacterial gene promoter lactose repressor (LacIpromoter), with the following ratio of components (wt.%): 0.0005–0.003 of interferon gamma with a specific activity of at least 1.7×107 IU/mg and purity of at least 97%; 0.005–0.01 of Twin 20; 2–8 of osmotic pressure regulators; 0.05–0.1 of buffer system maintaining pH 4.0–6.5; the rest is water.
EFFECT: invention makes it possible to create a liquid product for parenteral administration, which is a solution containing a purer chemically and biologically stabilized interferon gamma protein, which increased the effectiveness of the action and reduced its side effects due to the optimally selected composition of the components and their quantity, and also made it possible to increase the shelf life of the medicinal product to 3 years at the temperatures from 2 to 25°C.
3 cl, 4 dwg, 4 tbl, 12 ex
