OBJECTIVE:A new ionexchange
chromatographic procedure has been developed to a highly purified factor VIII conc-entrate from
plasma
cryoprecipitate.METHODS:Solubilized cryoprecipitate,after adsorption on aluminium hydroxide,acid precipitation in low temperature and PEG precipiatation,was treated with 0.3% tri(nbutyl)phosphate and 1% Tween 80 at 25℃ for at least 8hour to inactivate lipidenveloped viruses.The fractions then loaded onto a column packed with DEAEFractogel TSK 650M and chromatographed.Most proteins and TNBPTween 80 flowed through the gel unretarded.FVIII:c,which bound to the gel,was eluted by increasing the ionic strengh, then was directly filtersterilized without ultrafiltration or addition of a protein stablizier.RESULTS:
Chromatographic recovery of FVIII was 70%90%,and had a specific activity more than 100IU·mg -1 ,corresponding to a purification factor of over 5000 from plasma.CONCLUSION: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.