Purification of coagulation factor VIII using chromatographic methods. Direct chromatography of plasma in anion exchange resins.
Coagulation factor VIII (FVIII) concentrates are used in the treatment of patients with Hemophilia A. Human FVIII was purified directly from plasma using anion exchange chromatography followed by gel filtration. Three Q-Sepharose resins were tested, resulting in 40% recovery of FVIII activity using Q-Sepharose XL resin, about 80% using Q-Sepharose Fast Flow and 70% using the Q-Sepharose Big Beads. The vitamin K-dependent coagulation factors co-eluted with FVIII from the anion exchange columns. In the second step of purification, when Sepharose 6FF was used, 70% of FVIII activity was recovered free from vitamin K-dependent factors.
Cheng
Elisabeth
E
Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
Jinzenji
Daniela
D
Lorthiois
Ana Paula Almeida Aranha
AP
de Carvalho
Roberta Rodrigues
RR
Tanaka-Azevedo
Anita Mitiko
AM
Raw
Isaías
I
Martins
Elizabeth Angélica Leme
EA
D000837:Anion Exchange Resins; D001709:Biotechnology / Q000379:methods*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods; D005169:Factor VIII / Q000302:isolation & purification*; D006801:Humans; D010949:Plasma / Q000737:chemistry*; D013678:Technology, Pharmaceutical / Q000379:methods*
D000837:Anion Exchange Resins; C078147:F8 protein, human; D005169:Factor VIII