title: Progress in large-scale purification of factor VIII/von Willebrand factor concentrates using ion-exchange chromatography.
abstract: OBJECTIVE
  We investigated and optimized the parameters of a chromatographic process suitable for industrial scale to obtain a highly purified factor VIII (FVIII)/von Willebrand factor (VWF) concentrate.
  
  
  METHODS
  Several chromatographic runs were performed on the same production intermediate using different anion-exchange supports. The best matrix was selected and the final product was characterized. Once the chromatographic medium was chosen, the other parameters were evaluated to obtain the highest purified product and to modulate the VWF content in the FVIII/VWF complex.
  
  
  RESULTS
  Fractogel EMD TMAE was the best support among those tested. It was the only one maintaining good results either with standard or double loading and flow rate conditions with respect to a typical industrial process. The chromatographic recovery of FVIII co-purified with VWF was at least 86% with a specific activity not lower than 140 IU/mg. The FVIII/VWF complex obtained is highly pure and, with the exception of immunoglobulin M (IgM), all investigated contaminant proteins are under the detection limit. Different concentrates characterized by variable FVIII/VWF ratios were purified by varying the chromatographic conditions.
  
  
  CONCLUSIONS
  Several highly purified products, suitable for haemophilia A and von Willebrand disease management, can be obtained, through the same chromatographic process, on an industrial scale.
authors.affiliation: Kedrion Research Centre, Via Giovanni Pascoli 21, Castelvecchio Pascoli, Lucca, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Mori
authors.forename: I
authors.initials: I
authors.lastname: Nardini
authors.forename: P
authors.initials: P
authors.lastname: Rossi
authors.forename: C
authors.initials: C
authors.lastname: Nardini
authors.forename: C
authors.initials: C
authors.lastname: Farina
mesh_terms: D001781:Blood Component Removal / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004345:Drug Industry / Q000379:methods; D005169:Factor VIII / Q000302:isolation & purification*; D006801:Humans; D014841:von Willebrand Factor / Q000302:isolation & purification*
chemical_list: D014841:von Willebrand Factor; D005169:Factor VIII