NR AREQ
AU Geue,H.; Weber,P.; Kramer,K.; Giese,A.; Kretzschmar,H.A.; Hock,B.
TI Development of specific recombinant anti-PrPsc antibodies
QU TSE-Forum, 4. Kongress - Nationale TSE-Forschungsplattform, Düsseldorf 28.10.-29.10.2004, Poster DIA-03
PT Konferenz-Poster
AB
The goal of this project, which is part of the Bavarian Research Cooperation Prions (TUM 16), is the development of highly specific antibodies against different subtypes and isoforms of the prion protein. In this approach genetically engineered PrPsc-selective recombinant antibodies are aspired. These binders can be used as diagnostic tools for the detection of PrPsc.
PRNP knock-out mice are immunised with purified PrPsc from human Creutzfeldt-Jakob brains (cf. poster by Petra Weber et al.: Optimierung von Prion rod-Präparationen). Subsequent production of murine anti-PrPsc antibodies is monitored by ELISA techniques. mRNA is isolated from murine spleen and an antibody phage library is developed. For this purpose cDNA of the antibody encoding mRNA is acquired by RT-PCR. VH an VL genes are amplified with specific primers and ligated into a plasmid vector. Phages are produced in Escherichia coli. As the phages contain the genetic information for antibodies within their genome and present the corresponding antibody-fragment (single chain) on their surface, phages with specific antibody binding abilities can be selected by affinity chromatography. Subsequently, an evolutionary optimisation is performed by selecting phage antibodies with high affinity. The selected phages can further be genetically modified to improve the specificity of the corresponding antibody be repeated selection and exclusion.
AD Holger Geue, Karl Kramer, Bertold Hock, Technical University Munich, Chair of Cell Biology, Alte Akademie 12, D-85354 Freising; Petra Weber, Armin Giese, Hans Kretzschmar, Center for Neuropathology and Prion Research, Feodor-Lynen-Strasse 23, D-81377 Muenchen
SP englisch
PO Deutschland
OR Tagungsband