NR ARET
AU Hoffmann,T.; Ruoff,N.; Günter,J.; Klein,M.A.; Flechsig,E.
TI Characterization of novel monoclonal antibodies against PrP for diagnostic and therapeutic approaches
QU TSE-Forum, 4. Kongress - Nationale TSE-Forschungsplattform, Düsseldorf 28.10.-29.10.2004, Poster THE-03
PT Konferenz-Poster
AB
Antibodies against the prion protein (PrP) have become a powerful tool for diagnostic and therapeutic strategies in prion research. Recent studies have shown that antibodies inhibit prion propagation in cultured cells and prevent the onset of the disease in mice. Antibodies reacting with epitopes comprising residues 96-104 and 133-158 of PrPc possessed a particular potential inhibitory effect. However, an antibody that binds to the first epitope was found to trigger rapid and extensive apoptosis of neurons when applied to the brain. These findings encourage the belief that immunization protocols targeting the second epitope may provide a therapeutic approach to prion disease.
We have generated a panel of antibodies by immunizing PrP null mice with recombinant mouse PrP (recPrP) or its fibrillar form. Resulting hybridomas were screened by ELISA against recPrP and antibodies of stable clones were purified by protein G column. All 8 antibodies detect both PrPc and PrPsc in mouse brain homogenates by Western blotting and bind PrPc on the surface of murine N2A cells by FACS analysis. Four of them bind to PrPc on N2A cells with similar intensity as the commercial antibody 6H4. The binding affinity is approximately 2- to 10-fold lower than that of 6H4. The epitopes of 7 antibodies were located between residues 141 and 176. Only one antibody (B2-31-77) binds to the region comprising residues 93 to 106. Furthermore, we examined the ability of three antibodies to inhibit prion propagation in RML-infected N2A cells. The levels of PrPsc in cells that were cultured in the presence of the tested antibody were strongly diminished in comparison to that in mock-treated cells or in cells cultured with an antibody against the neural cell-adhesion molecule (NCAM). These PrP specific antibodies will be further analyzed and used for diagnostic purposes and therapeutic approaches in mouse models.
AD Tanja Hoffmann, Nele Ruoff, Julia Günter, Michael A. Klein and Eckhard Flechsig, Prion Research Group, Institute of Virology and Immunobiology, University of Würzburg
SP englisch
PO Deutschland
OR Tagungsband