METHOD OF ANTITROMBIN III MEDICATION PREPARATION OF HUMAN PLASMA Russian patent published in 2009 - IPC A61K35/14 A61K38/16 A61K38/55 A61P7/02 

FIELD: pharmacology.

SUBSTANCE: invention relates to the fields of biotechnology and pharmaceutical industry immediately dealing with preparation of Antitrombin III medication of human plasma. The method proposed envisages such operations as affinity chromatography, decontamination of viruses and product concentration. Plasma is pre-centrifuged. The sediment detached, PEG 4000 is added to the supernatant. Then follow the operations of incubation, centrifugation, diafiltration against a pH 7.20 buffer solution containing tris-HCl and sodium citrate and flowing through a DEAE-sepharose column. Tri-n-butylphosphate and Twin-80 are added to the plasma protein mixture having failed to become bound to the medium. The mixture undergoes incubation and is applied onto a Q-sepharose column. The protein fraction containing the target protein becomes eluted with a pH 9.0 tris-HCl buffer with a conductance of 40.0±0.8 mSi. The eluate becomes dialysed against a pH 7.40 tris-HCl buffer containing sodium chloride and sodium citrate and pasteurised at 60°C during 10 hours. The ATIII-enriched protein mixture becomes dialysed against a pH 7.40 buffer containing tris-HCl and sodium citrate and applied onto a heparin-sepharose column. The column is washed with the same buffer additionally containing sodium chloride. The target product becomes eluted with the same buffer additionally containing 1.50±0.03 M sodium chloride. The ATIII solution becomes diafiltered against a formulating buffer, sterilely filtered, dispensed and lyophilically dried.

EFFECT: product yield improvement in terms of activity by a maximum of 60% as well as provision for the product high quality and pureness.

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