A sensitive filter retention assay for the detection of PrP(Sc) and the screening of anti-prion compounds.
A hallmark of prion diseases is the accumulation of an abnormally folded prion protein, denoted PrP(Sc). Here we describe a new and highly sensitive method for the detection of PrP(Sc) in brain and other tissue samples that utilizes both PrP(Sc) diagnostic criteria in combination; protease resistance and aggregation. Upon filtration of tissue extracts derived from scrapie- or bovine spongiform encephalopathy-infected animals, PrP(Sc) is retained and detected on the membranes. Laborious steps such as SDS-PAGE and Western blotting are avoided with concomitant gain in sensitivity and reliability. The new procedure also proved useful in a screen for anti-prion compounds in a scrapie-infected cell culture model.
Winklhofer
K F
KF
Department of Cellular Biochemistry, Max-Planck-Institut für Biochemie, D-82152, Martinsried, Germany.
Hartl
F U
FU
Tatzelt
J
J
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002417:Cattle; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D016643:Encephalopathy, Bovine Spongiform / Q000378:metabolism; D051379:Mice; D018620:PrPSc Proteins / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D018620:PrPSc Proteins