Application of dye-ligand chromatography for purification of coagulation factor VIII

Abstract

The problem of processing of donor plasma for creating an effective medications is important. Nowadays a big number of methods for fractionation of plasma separation and purification components, especially coagulation factors and fibrinolysis were described. The methods of biospecifical chromatography are especially attractive. The chromatographic purification of factor VIII is a trouble due to potential risks of activation and instability in the presence of plasma proteases. The anion exchange, gel-filtration and affinity chromatography are widely used for purification of coagulation factor VIII. The synthetic active triazine dyes have a number of advantages as ligands. We explored the possibility of using dye-ligand affinity chromatography on silica carriers for the isolation and purification of clotting factor VIII. The results of studies of sorption this factor with different chromatographic sorbents and the method of its purification from cryoprecipitate by binding of additional proteins on Diasorb aminopropyl with immobilized triazine active dyes are described. As a result of this work, we have selected a group of sorbents are the best for purification of factor VIII of blood coagulation: Diasorb-Active scarlet damask 4GT, Diasorb-Procion gelb M4R, Diasorb-Procion blue HB, Diasorb-Procion blue MXR and Diasorb-Active bright blue K. The purification of factor VIII occured due to the phenomenon of negative affinity chromatography. The specific activity of factor increased approximately tenfold.

Keywords

• blood coagulation factor VIII
• affinity chromatography
• triazine active dyes