Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 84, Issue 1, Pages 77-89Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0166-0934(99)00135-4
Keywords
Western blot assay; transmissible spongiform encephalopathy; prion protein
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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 PrPRES, 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 PrPRES down to similar to 10(3.4) IU/ml, which is similar to 5-10 pg of PrP or similar to 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. (C) 2000 Elsevier Science B.V. All rights reserved.
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