NR ATOG
AU Hoffmann,T.; Ruoff,N.; Guenther,J.; Klein,M.A.; Flechsig,E.
TI Characterization of novel monoclonal antibodies against PrP
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Human prions, risk of blood products, and therapy HUMAN-30
PT Konferenz-Poster
AB Antibodies against the prion protein (PrP) have become a powerful tool for therapeutic strategies in prion research. Antibodies reacting with epitopes comprising residues 96-104 and 133-158 of PrPc possessed a particular potential inhibitory effect in vitro and in vivo. 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 or its fibrillar form. Resulting hybridomas were screened by ELISA and antibodies of stable clones were purified by protein G column. All antibodies analysed so far 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. Five of them bind to PrPc on N2A cells with similar intensity as the commercial antibody 6H4. The epitopes of 6 antibodies were located between residues 141 and 176. Only one antibody (B2-31-77) binds to the region comprising residues 93 to 106. Four of six antibodies tested were able to inhibit prion propagation in RML-infected N2A cells. The PrP-specific antibodies will be further analyzed and used for diagnostic purposes and therapeutic approaches in mouse models.
AD T.Hoffmann, N.Ruoff, J.Guenther, M.A.Klein, E.Flechsig, University Wuerzburg, Institute of Virology and Immunobiology
SP englisch
PO Deutschland