METHOD FOR PRODUCING LIQUID IMMUNOGLOBULIN AGAINST EBOLA FEVER FROM HORSE BLOOD SERUM Russian patent published in 2018 - IPC A61K39/42 C07K1/16 C07K1/18 C07K16/06 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, namely to development of technologies for production of protective immunobiological preparations, and concerns the preparation of Ebola immunoglobulin from horses' serum, immunized with the Ebola virus, by the Kohn alcohol precipitation method. Method for obtaining a heterologous immunoglobulin against Ebola fever includes additional purification and concentration steps of the preparation using ion exchange and gel filtration chromatography methods, immunoglobulin against Ebola fever, including ion-exchange chromatography on a column BPG 140/500 GE Health care with QSepharose FF at a temperature of 20 °C and a sample feed rate of 350 ml/min followed by gradient elution of the Ig G fraction from 0 to 100 % with a buffer solution with a pH of 8.0 containing 0.02 mol/l tris(hydroxy-methyl)aminomethane and 1 mol/l NaCl at the same rate for 80 minutes; gel filtration chromatography on a BPG 140/950 GE Health care column with 40 SEC sorbent, equilibrated with a phosphate-glycine buffer solution with pH 6.9 containing 0.02 M sodium phosphate with 2 % glycine at a flow rate of 100 ml/min in an isocratic mode of elution at the same rate; concentration and sterilizing filtration of the immunoglobulin solution on an automatic filtration system and concentration in the tangential flow “Vladisart” through cassettes with exclusion limit of 100 kDa at a flow rate of 500 ml/min, an inlet pressure of not more than 2 bar and an outlet pressure of not more than 1 bar.

EFFECT: technical result consists in that the developed method provides immunoglobulin against Ebola fever from the blood serum of horses with a higher content of monomeric immunoglobulin fraction (not less than 95 %) and a smaller number of extraneous fractions of dimers, polymers and aggregates (no more than 5 %), as well as the absence of residual ethyl alcohol, which reduces the anaphylactogenicity of immunoglobulin and leads the quality of the drug in accordance with the requirements of the European Pharmacopeia for heterologous drugs.

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