Purification of recombinant human coagulation factors II and IX and protein S expressed in recombinant Vaccinia virus-infected Vero cells.
Recombinant human coagulation factors II and IX and recombinant human protein S were expressed by the Vaccinia virus expression system in Vero cells and isolated from the serum-free media. All three recombinant proteins were purified by the same universal strategy. After concentration by anion exchange chromatography, recombinant coagulation factors were purified by anion exchange filtration in the presence of Ca2+. The addition of Ca2+ to the vitamin K-dependent coagulation factors modified their physico-chemical behavior during anion exchange chromatography and resulted in the separation of the recombinant coagulation proteins from contaminating proteins.
Fischer
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B
IMMUNO AG, Biomedical Research Center, Donau, Austria.
Mitterer
A
A
Dorner
F
F
D000818:Animals; D001777:Blood Coagulation; D002118:Calcium; D002522:Chlorocebus aethiops; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D016895:Culture Media, Serum-Free; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005164:Factor IX / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D005822:Genetic Vectors; D006801:Humans; D017293:Protein S / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D011516:Prothrombin / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D014162:Transfection / Q000379:methods; D014616:Vaccinia virus; D014709:Vero Cells
D016895:Culture Media, Serum-Free; D017293:Protein S; D011994:Recombinant Proteins; D011516:Prothrombin; D005164:Factor IX; D002118:Calcium