Monitoring plasma processing steps with a sensitive Western blot assay forthe detection of the prion protein

Citation
Dc. Lee et al., Monitoring plasma processing steps with a sensitive Western blot assay forthe detection of the prion protein, J VIROL MET, 84(1), 2000, pp. 77-89
Citations number
40
Categorie Soggetti
Microbiology
Journal title
JOURNAL OF VIROLOGICAL METHODS
ISSN journal
01660934 → ACNP
Volume
84
Issue
1
Year of publication
2000
Pages
77 - 89
Database
ISI
SICI code
0166-0934(200001)84:1<77:MPPSWA>2.0.ZU;2-3
Abstract
Determining the risk of transmissible spongiform encephalopathy (TSE) trans mission by blood or plasma-derived products requires sensitive and specific assays for the detection of either infectivity or a reliable marker for in fectivity. To this end, a Western blot assay that is both sensitive and rep roducible for the detection of PrPRES, a marker for TSE infectivity, was de veloped. Using the 263K strain of TSE as a model system, the Western blot a ssay proved to be sensitive, specific and quantitative over a 3-4 log dynam ic range. Compared to the rodent bioassay, the assay was shown to detect Pr PRES down to similar to 10(3.4) IU/ml, which is similar to 5-10 pg of PrP o r similar to 10-20 ng brain equivalents. The Western blot was applied to mo nitor the partitioning of spiked PrPSc through three plasma fractionation s teps, cryoprecipitation, fraction I and Fraction III, that are common to th e purification of several human plasma-derived therapeutic products includi ng albumin and immunoglobulins. The results from these studies demonstrated 1 log, 1 log and 4 logs of PrPSc partitioning away from the effluent fract ion for the cryoprecipitation, fraction I and fraction III steps, respectiv ely. (C) 2000 Elsevier Science B.V. All rights reserved.