NR AOSZ
AU Geue,H.; Seifert,M.; Hock,B.
TI Plasminogen-Prion Binding Studies
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-39
PT Konferenz-Poster
AB
Plasminogen (Plg) has been reported to bind specifically to infectious prion proteins (PrPsc) under high detergent concentration (Fischer et al. 2000). In our study we try to utilize affinity for the production of monoclonal PrPsc-specific antibodies for analytical and therapeutical applications. Plg is used as immunogen to produce anti-Plg antibodies whose binding sites are congruent with PrPsc. In a second step rabbits are immunised with fragments of these antibodies, producing anti-idiotypic antibodies, which are congruent with Plg and therefore bind PrPsc.
Mice were immunised with the Plg fragment LBS I (Lysin binding site 1) and an adaption of the published Western Blot method to ELISA applications was made. ELISAs (several formats) with Plg yielded no significant affinity for PrPsc. Therefore we investigated the postulated Plg-PrPsc interaction in Western Blot experiments. Homogenised infectious material from bovine and hamster brain from different sources were applied. No significant interaction between Plg and PrPsc was detected - neither following the original procedure nor with optimised protocols.
Since the production of anti-PrPsc monoclonal antibodies requires a high Plg-PrPsc binding affinity, the very weak affinity found in our experiments does not allow a development of specific anti-PrPsc antibodies or their application for ELISA or other diagnostic tools. Alternatives are discussed.
AD Holger Geue, Martin Seifert, Bertold Hock, Technische Universität Muenchen, Germany
SP englisch
PO Deutschland