FIELD: chemistry.
SUBSTANCE: method of purifying coagulation factor VIII protein from a solution involves: a) contacting the protein with a multimodal resin or a mixed action-type resin containing ligands having a hydrophobic portion and a negatively charged portion; b) elution of the protein with an elution buffer containing at least 1.5M salt and at least 40% (wt/vol) ethylene glycol, propylene glycol or mixture thereof, and calcium ions. The method is a single-step chromatographic process where the coagulation factor VIII protein is captured and does not require adjusting pH or electroconductivity at the feeding step. The method of stabilising the coagulation factor VIII protein, where the coagulation factor VIII protein is captured and stabilised when the protein passes once through a column with a multimodal resin containing ligands having a hydrophobic portion and a negatively charged portion, and elution of the protein with an elution buffer containing at least 1.5M salt and at least 40% (wt/vol) ethylene glycol, propylene glycol or mixture thereof, and calcium ions.
EFFECT: invention reduces the volume of the column about 250-fold and a coefficient of purification equal to 30, high stability of the protein product.
2 cl, 4 dwg, 4 tbl, 7 ex
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Authors
Dates
2013-09-20—Published
2008-07-11—Filed