Purification of factor VIII:c coagulant activity from rat liver nonparenchymal cell culture medium by immobilized metal ion affinity chromatography.
The purification of factor VIII:c coagulant activity on the basis of its affinity for calcium is described. For this purpose, use was made of a recently introduced chelating matrix, i.e., carboxymethylated aspartic acid agarose, coupled with calcium--thereby creating a gel with specificity comparable with biospecific affinity chromatography. In a single step factor VIII:c activity was purified from rat liver nonparenchymal cell culture medium with a purification factor of 85-fold. The material exhibits a single band on polyacrylamide gel electrophoresis.
Mantovaara
T
T
Institute of Biochemistry, Uppsala University, Sweden.
Pertoft
H
H
Porath
J
J
D000818:Animals; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005169:Factor VIII / Q000302:isolation & purification*; D008099:Liver / Q000737:chemistry* / Q000166:cytology; D008297:Male; D008670:Metals; D051381:Rats; D011919:Rats, Inbred Strains
D003470:Culture Media; D008670:Metals; D005169:Factor VIII