NR ATLR
AU Geue,H.; Weber,P.; Kramer,K.; Giese,A.; Kretzschmar,H.A.; Hock,B.
TI Development of specific recombinant anti-PrPsc antibodies
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-55
PT Konferenz-Poster
AB
The goal of this project 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. Stimulation 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 obtained by RT-PCR. VH and VL antibody genes are amplified with specific primers and inserted into a phagemid vector. Recombinant phages are produced in Escherichia coli. As the phages contain the genetic information for antibodies within their genome and display the corresponding antibody fragment (single-chain) on their surface, phages with specific antibody binding properties are selected by affinity chromatography. The selected phages can further be genetically modified to improve the affinity and specificity by reiterative evolutionary cycles.
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 A. Kretzschmar, Ludwig-Maximilans-University Munich, Centre of Neuropathology and Prion Research, Feoodr-Lynen-Str. 23, D-81377 Munich
SP englisch
PO Deutschland