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Removal of prion challenge from an immune globulin preparation by use of a size‐exclusion filter
Author(s) -
Van Holten Robert W.,
Autenrieth Stephen,
Boose Jeri Ann,
Hsieh W.T.,
Dolan Sherri
Publication year - 2002
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1046/j.1537-2995.2002.00160.x
Subject(s) - ultrafiltration (renal) , sonication , western blot , filtration (mathematics) , chromatography , size exclusion chromatography , chemistry , globulin , immune system , scrapie , centrifugation , antibody , biology , microbiology and biotechnology , biochemistry , prion protein , immunology , medicine , statistics , mathematics , disease , gene , enzyme
BACKGROUND : A size‐exclusion filter (Viresolve 180, Millipore Corp.) was tested for its ability to remove transmissible spongiform encephalopathies prion protein from an immune globulin preparation during ultrafiltration. STUDY DESIGN AND METHODS: Hamster‐adapted 263K scrapie brain homogenate (SBH) was spiked into Rho(D) immune globulin (human) at 1 in 300 and 1 in 1000 dilutions. Before spiking, the SBH was treated with detergent, sonicated, and filtered through serial 0.45‐, 0.22‐, and 0.1‐μm filters to present a rigorous filter challenge. Process variables were monitored throughout the ultrafiltration to ensure that the spiked material did not compromise the membrane flux. Removal of scrapie prion protein (PrPSc ) material was determined by use of a sensitive Western blot assay. RESULTS: The turbid SBH became completely translucent after sonication and passage through the 0.45‐, 0.22‐, and 0.1‐μm filters. The filtration of the immune globulin containing PrPScmaterial was more difficult to perform than was filtration of immune globulin spiked with the normal cellular isoform. Even during tangential flow filtration, the fibril material prevented the PrPSc‐spiked immune globulin from passing as readily through the filter. Western blot results indicated a removal of greater than or equal to 2.5 log PrPSc , while remaining within the normal filtration limits. CONCLUSIONS: The composition, physical condition, and the amount of SBH introduced have significant effects on the filtration of the immune globulin and the log removal values obtained. By use of a detergent‐treated, sonicated, and filtered preparation of SBH, it was demonstrated that the Viresolve 180 effectively removes PrPSc from the immune globulin.