Premium
Transmissible spongiform encephalopathy agent clearance by the immunoaffinity and anion‐exchange chromatography steps of the R e F acto ® manufacturing process
Author(s) -
BOOTH J.,
VICIK S.,
TANNATT M.,
GALLO C.,
KELLEY B.
Publication year - 2007
Publication title -
haemophilia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.213
H-Index - 92
eISSN - 1365-2516
pISSN - 1351-8216
DOI - 10.1111/j.1365-2516.2007.01441.x
Subject(s) - sepharose , recombinant dna , ion chromatography , affinity chromatography , chromatography , hamster , chemistry , virology , biochemistry , medicine , microbiology and biotechnology , biology , enzyme , gene
Summary. R e F acto ® (moroctocog alfa), a recombinant factor VIII approved for the treatment of haemophilia A, is produced by a mammalian cell‐culture process that includes therapeutic‐grade human serum albumin (HSA) in the cell‐culture medium. While to date there have been no cases of transmissible spongiform encephalopathy (TSE) resulting from the clinical use of HSA, Wyeth conducted a study to demonstrate that the R e F acto manufacturing process has significant capacity to remove a TSE agent if it were present as a contaminant in the HSA. The immunoaffinity (8A4 Sepharose) and anion‐exchange (Q Sepharose) chromatography steps were evaluated for the clearance of the hamster TSE agent, strain 263K. This Good Laboratory Practice study was performed using appropriately qualified, laboratory‐scale chromatography systems. Filtered brain homogenate from TSE‐infected hamsters was added to loads of both chromatographic columns, and the concentration of TSE agent in the loads and product pools were determined using a validated western blot quantitation method. Replicate chromatography runs were consistent, as demonstrated by the ≤0.7 log 10 difference observed in TSE agent reduction between each pair of runs. The immunoaffinity and anion‐exchanges steps demonstrated 3.8 log reduction and >5.2 log reduction respectively. These data provide a high degree of assurance that in the unlikely event of a TSE contamination of the HSA used in the R e F acto cell‐culture process, the purification steps have the potential to remove the infectious agent to extremely low levels, thereby significantly reducing the risk to patients receiving R e F acto .