Author/Editor     Josić, Djuro; Schwinn, Horst; Stadler, M; Štrancar, Aleš
Title     Purification of factor VIII and von Willebrand factor from human plasma by anion-exchange chromatography
Type     članek
Source     J Chromatogr B Biomed Appl
Vol. and No.     Letnik 662, št. 2
Publication year     1994
Volume     str. 181-90
Language     eng
Abstract     Factor VIII (anti-hemophilia A factor) is isolated from human plasma. Purification is carried out by a combination of precipitation and chromatographic procedures. After precipitation, the first step in virus inactivation is achieved through the effect of a non-ionic detergent such as Tween 80, and a solvent, e.g. tri-n-butylphosphate (TnBP). By subsequent anion-exchange chromatography, a highly enriched product is isolated, consisting of a complex formed by factor VIII and von Willebrand factor (FVIII-vWF). This treatment also removes the virus-inactivating reagents to quantities in the low ppm range. The second step in virus inactivation is aimed specifically at the non-enveloped viruses and consists of pasteurization at temperatures higher than 60 degrees C for 10 h. Through the addition of stabilizers, between 80% and 90% of the initial activity of FVIII is preserved during the modified pasteurisation. Along with the possibly denatured proteins the stabilizers, such as sugars, amino acids and bivalent cations, are subsequently removed by ion-exchange chromatography. The two-fold virus inactivation, by solvent/detergent treatment and subsequent pasteurisation, allows the destruction of both lipid-enveloped and non-enveloped viruses. During the procedure FVIII is stabilized through the high content of vWF. The complex consisting of FVIII and vWF can be dissociated by adding calcium ions. Subsequently both glycoproteins from this complex are separated from one another by further anion-exchange chromatography.
Descriptors     FACTOR VIII
VON WILLEBRAND FACTOR
VIRUSES
ULTRAFILTRATION
QUALITY CONTROL
PLASMA
ION EXCHANGE RESINS
ELECTROPHORESIS, POLYACRYLAMIDE GEL
CHROMATOGRAPHY, ION EXCHANGE
BIOCOMPATIBLE MATERIALS