NR AOWS
AU Mehl,M.; Kuhn,F.; Miglino,N.; Fischer,K.; Raeber,A.J.; Oesch,B.
TI Characterisation of normal prion protein in bovine and mouse urine
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-102
PT Konferenz-Poster
AB Urine represents an attractive matrix for a diagnostic test. We therefore have undertaken to characterize normal PrP from urine as well as the conditions to optimally detect it using our current diagnostic tests (i.e. Prionics Check Western blot as well as sandwich immunoassays. It proved to be difficult to directly detect PrP in urine; we have therefore used a size exclusion filtration to concentrate the prion protein and further purify it by anion exchange or immunoaffinity chromatography. Labelled anti-PrP antibodies detect a smeared PrP band in the molecular range of 20-30 kD but in some cases the signal was very diffuse probably due to size heterogeneity. Using sandwich immunoassays, PrP from urine could be readily detected suggesting that the central portion of PrP is present abundantly. Assays were optimized using urine from knockout, wild type and PrP overexpressing mice allowing discrimination between real signals due to PrP and non-specific signals. Furthermore, we found that the sampling procedure and timing is crucial to achieve reproducible results. We intend to further characterise the different forms of PrP from urine and apply our detection know how to samples from infected animals.
AD Martin Mehl, Franziska Kuhn, Nicola Miglino, Karin Fischer, Alex Raeber, Bruno Oesch, Prionics AG, Switzerland
SP englisch
PO Deutschland