Abstract Number: PB0490
Meeting: ISTH 2021 Congress
Background: The factor VIII (FVIII) concentrate should have to high specific activity, viral security, stability and don’t cause the appearance of anti-FVIII antibodies. It’s known that the use of FVIII concentrate with a high content of factor von Willebrande (vWF) reduces the risk of development of anti-FVIII antibodies.
Aims: to investigate and describe the main properties of the FVIII concentrate.
Methods: The Fresh Frozen Plasma was used as initial material. The activity of FVIII was determined using a coagulation test that measures the partial thromboplastin time; vWF – the cofactor activity of rystomycin; the total concentration of proteins – the Bradford method; the albumine – with bromcresol green. The ion-exchange and dye-ligand affinity chromatography have been performed.
Results: We have been developed a method of purification of FVIII by the methods ion-exchange and dye-ligand affinity chromatography. The obtained concentrate of FVIII contain vWF (3,07±0,18 IU/ml) and albumine (0,67±0,02×10-1 mg/ml). The both of these proteins are used as stabilizers of its activity. It’s known that some researchers suggest using of vWF as a natural stabilizer of FVIII activity and doesn’t require the addition of stabilizers. In our case, the resulting drug additionally contains these two proteins. The researches of the stability of FVIII concentrate have been demonstrated the following: the activity of FVIII in solution is reduced by 20% after 6–8 hours, by 40% – after 24 hours and by 90% after 48 hours at room temperature. The activity of the freezing factor is fully preserved during the month, after three months of storage it decreases by 10%, after a year of storage – by 20–25%. Lyophilization of the concentrate ensures the preservation of its activity for two years at +(4÷8) ºC.
Conclusions: We have been developed a new method purification of FVIII which doesn’t require the additional proteins as stabilizers.
To cite this abstract in AMA style:Shurko N, Danysh T, Novak V. Properties of FVIII Concentrate Were Obtained by the Method of Dye-ligand Affinity Chromatography [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/properties-of-fviii-concentrate-were-obtained-by-the-method-of-dye-ligand-affinity-chromatography/. Accessed October 1, 2023.
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