Large-scale gel filtration chromatography for the production of a solvent/detergent-treated high-purity factor VIII concentrate.
To produce a tri(n-butyl)phosphate/sodium-cholate-treated intermediate-purity factor VIII (FVIII) concentrate with a specific activity of about 1 IU/mg, we used a simple gel filtration step with Sephadex G25 to remove the solvent/detergent reagents from the final product. By exchanging the Sephadex G25 gel for a new high-resolution gel (Sephacryl S400 HR), we obtained a high-purity FVIII concentrate, by simultaneous elimination of about 98% of the extraneous proteins and removal of the solvent/detergent reagents, without reducing the FVIII:c yield and without altering the production scheme. With different protein analysis techniques we analysed the resulting FVIII concentrate and, comparing it to the formerly produced intermediate-purity FVIII concentrate, demonstrated improved purity.
DRK Blutspendezentrale, Baden-Baden, BRD.
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Dengler
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Stöcker
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Kellner
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Fürst
D019826:Cholic Acid; D002793:Cholic Acids / Q000032:analysis*; D002850:Chromatography, Gel; D003902:Detergents; D003911:Dextrans; D005169:Factor VIII / Q000302:isolation & purification*; D005782:Gels; D006801:Humans; D010755:Organophosphates / Q000032:analysis*; D009943:Organophosphorus Compounds / Q000032:analysis*; D012997:Solvents
D002793:Cholic Acids; D003902:Detergents; D003911:Dextrans; D005782:Gels; D010755:Organophosphates; D009943:Organophosphorus Compounds; D012997:Solvents; D005169:Factor VIII; C025614:sephadex; C009524:tributyl phosphate; D019826:Cholic Acid