Development, upscaling and validation of the purification process for human-cl rhFVIII (Nuwiq®), a new generation recombinant factor VIII produced in a human cell-line

摘要: Abstract Introduction Human-cl rhFVIII (Nuwiq®), a new generation recombinant factor VIII (rFVIII), is the first rFVIII produced in human cell-line approved by European Medicines Agency. Aims To describe development, upscaling and process validation for industrial-scale human-cl purification. Methods results The purification involves one centrifugation, two filtration, five chromatography columns dedicated pathogen clearance steps (solvent/detergent treatment 20 nm nanofiltration). key step uses an affinity resin (VIIISelect) with high specificity FVIII, removing essentially all host-cell proteins >80% product recovery. production-scale multi-step efficiently removes process- product-related impurities high-purity product, overall yield of ∼50%. Specific activity final was >9000 IU/mg, ratio between active FVIII total protein present >0.9. entire production free animal-derived products. Leaching potential harmful compounds from resins pathogens tested were below limit quantification product. Conclusions can be at 500 L bioreactor scale, maintaining purity recoveries. innovative ensures high-quality safety margin.