FIELD: chemistry.
SUBSTANCE: disclosed is a method of producing, separating and purifying the human granulocyte colony stimulating factor (hG-CSF). Cells of a recombinant producer strain are obtained and the culture fluid is then concentrated. Bacterial cells are broken down while separating inclusion bodies via double treatment with high pressure. Protein impurities are removed. The inclusion bodies are dissolved and protein is recovered. Protein renaturation is carried out for 70-90 hours while diluting the protein solution to 400 ml. The obtained solution is deposited on an anion-exchange chromatographic column with diameter of 300 mm with linear rate of deposition of 600-1000 ml/min. The hG-CSF is then concentrated successively on chromatographic columns with anion-exchange or cation-exchange sorbents. The hG-CSF is then purified on a cation exchanger in fraction recycling conditions and stabilised by dialysis.
EFFECT: method enables to obtain, in one cycle, a stable preparation of hG-CSF in amount of 15-20 g with high homogeneity on HPLC and high hemostimulating activity.
5 cl, 1 dwg, 2 ex
