Premium
A Highly Purified Factor VIII:c Concentrate Prepared from Cryoprecipitate by Ion‐Exchange Chromatography
Author(s) -
Burnouf T.,
BurnoufRadosevich M.,
Huart J. J.,
Goudemand M.
Publication year - 1991
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.1991.tb00864.x
Subject(s) - cryoprecipitate , chemistry , chromatography , immunoadsorption , elution , ion chromatography , ionic strength , ultrafiltration (renal) , fibrinogen , antibody , biochemistry , aqueous solution , immunology , biology
. A new ion‐exchange chromatographic procedure has been developed to produce a highly purified factor VIII (FVIII) concentrate from plasma cryoprecipitate. Solubilized cryoprecipitate, after adsorption on aluminium hydroxide and cold precipitation, was treated with 0.3% tri(n‐butyl)phosphate and 1% Tween 80 at 25°c for at least 8 h to inactivate lipid‐enveloped viruses. The fraction was then loaded onto a column packed with DEAE‐Fractogel TSK 650 M and chromatographed. Most proteins and TnBP‐Tween 80 flowed through the gel unretarded. FVIII:c, which bound to the gel, was eluted by increasing the ionic strength, then was directly filter‐sterilized without ultrafiltration or addition of a protein stabilizer. Chromatographic recovery of FVIII:c was 80–90%. After freeze‐drying, FVIII:c was at a concentration of 42.5 ± 9.5 IU/ml and had a specific activity of 175.4 ± 37.8 IU/mg (n = 40), corresponding to a purification factor of over 12,000 from plasma. The typical yield of the freeze‐dried FVIII:c from cryoprecipitate was 55–65%. FVIII:c was stable for over 24 h at room temperature in the liquid state. The mean content of fibrinogen and immunoglobulin G was only 65 and 100mg/l, respectively, corresponding to 1.4 and 2.3 mg/1,000 IU FVIII:c. This concentrate, which is much purer than traditional FVIII concentrates, has been found to be well tolerated and effective in clinical treatment of hemophilia A patients.