NR AHDH
AU Lee,D.C.; Stenland,C.J.; Hartwell,R.C.; Ford,E.K.; Cai,K.; Miller,J.L.; Gilligan,K.J.; Rubenstein,R.; Fournel,M.; Petteway,S.R.Jr.
TI Monitoring plasma processing steps with a sensitive Western blot assay for the detection of the prion protein
QU Journal of Virological Methods 2000 Jan; 84(1): 77-89
PT journal article
AB Determining the risk of transmissible spongiform encephalopathy (TSE) transmission by blood or plasma-derived products requires sensitive and specific assays for the detection of either infectivity or a reliable marker for infectivity. To this end, a Western blot assay that is both sensitive and reproducible for the detection of PrP(RES), a marker for TSE infectivity, was developed. Using the 263K strain of TSE as a model system, the Western blot assay proved to be sensitive, specific and quantitative over a 3-4 log dynamic range. Compared to the rodent bioassay, the assay was shown to detect PrP(RES) down to approximately 10(3.4) IU/ml which is approximately 5-10 pg of PrP or approximately 10-20 ng brain equivalents. The Western blot was applied to monitor the partitioning of spiked PrPsc through three plasma fractionation steps, cryoprecipitation, fraction I and fraction III, that are common to the purification of several human plasma-derived therapeutic products including albumin and immunoglobulins. The results from these studies demonstrated 1 log, 1 log and 4 logs of PrPsc partitioning away from the effluent fraction for the cryoprecipitation, fraction I and fraction III steps, respectively.
MH Amino Acid Sequence; Animal; Biological Assay; Blotting, Western/*methods/statistics & numerical data; Brain Chemistry; Comparative Study; Epitopes/genetics; Freezing; Hamsters; Human; PrPsc Proteins/blood/genetics/immunology; Precipitation; Prion Diseases/blood/transmission; Prions/*blood/genetics/immunology; Sensitivity and Specificity; Virology/*methods/statistics & numerical data
AD Department of Pathogen Safety Research/Biological Products, Bayer Corp., Research Triangle Park, NC 27709, USA. doug.lee.b@bayer.com
SP englisch
PO Niederlande