NR AXPB
AU Jackson,G.S.; Pal,S.; Jones,S.; Cooper,S.; Khalili-Shirazi,A.; Hawke,S.; Wadsworth,J.D.F.; Collinge,J.
TI Molecular Diagnosis of Human Prion Disease using an Antibody Specific for PrPsc
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.144
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: Disease-associated prion protein, PrPsc, is a reliable marker of prion disease and its detection has become a standard diagnostic method. A major limitation of this strategy is that amino acid sequence identity between the normal cellular form of the prion protein, PrPc, and PrPsc results in cross-reactivity in immunoassays. The low abundance of PrPsc against a background of ubiquitously expressed PrPc typically requires depletion of PrPc, usually by proteolytic digestion of the diagnostic specimen, limiting the sensitivity of diagnostic assays. It is also now apparent that protease-sensitive pathological isoforms of PrP may have a significant role in prion diseases and therefore new diagnostic tests that do not rely on proteolysis are required.
Methods: By immunisation with an altered conformation of the human prion protein, ß-
PrP, we have isolated a monoclonal antibody that exhibits selective binding to PrPsc. Protocols for the immunoprecipitation of PrPsc with this antibody coupled with western blot detection allow diagnosis of CJD without the use of proteolysis.
Results: Use of this novel antibody allows accurate diagnosis of vCJD with 100% sensitivity and specificity and detection of PrPsc in cases where standard western blotting fail to detect protease resistant PrP.
Discussion: We have isolated and characterised an antibody that is selective for disease-associated PrP and we describe a method for the diagnosis of CJD which does not require PrPc depletion by proteolysis or other methods. This antibody may facilitate the sensitive detection of PrPsc in peripheral tissues or fluids where the ratio of PrPc to PrPsc is high and proteolysis does not provide sufficient discrimination.
AD G.S. Jackson, S. Pal, S. Jones, S. Cooper, A. Khalili-Shirazi, J.D.F. Wadsworth, J. Collinge, Institute of Neurology, MRC Prion Unit, UK; S. Hawke, University of Sydney, The Medical Foundation, Australia
SP englisch
PO Schottland
EA pdf-Datei und Poster (Posterautoren: S. Pal, S. Jones, S. Cooper, A. Khalili, J.D.F. Wadsworth, J. Collinge und G.S. Jackson)