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Use of novel recombinant prion proteins to generate range of antibodies and peptides that selectively bind aminoterminus (follow on SE2009) - SE1765

Understanding the conformational state of prion protein is undoubtedly key to understanding the mechanism of TSE pathogenesis and diagnosis. As part of studies aimed at understanding the 3D structure of PrP,we have produced soluble high yielding sources of mouse prion protein and shown conformational changes in the amino terminus of PrP associated with the acquisition of copper (and other metal) ions. The copper-liganded form of PrP is different from many forms of non-copper bound PrP studied to date. Copper bound PrP has a distinct conformation as shown by CID spectra and antibody binding, exhibits a novel enzyme activity and shows an unusually high oxidation of methionine residues. Copper can be replaced with manganese to produce a molecule that has some features shared by PrPsc. As metal ion binding occurs to the repeat sequences in the amino terminus of PrP, metal and non-metal bound forms of PrP may be useful to select antibodies or peptides that specifically target this region.

As PrPSc conformational changes also occur in the amino terminal half of PrP, reagents purposefully selected to this region may have potential as discriminatory reagents. Thus, the main objective of this proposal is to use the copper and manganese liganded forms of PrP to select antibodies and peptides that recognise differences between the various PrP conformations.

The study will be enabled by a unique collaboration that uses our experience with recombinant PrP production and the unparalleled expertise of Cambridge Antibody Technology in phage antibody display systems. The proposed work addresses directly research requirement 1.1 of the TSE Research and Surveillance Unit research requirements document 2001/2002, specifically the recognition that Proposals aimed at understanding the structural differences between different isoforms of PrP may lead to differential diagnostic tests. The intended use of this work is in basic PrP research, where antibodies and peptides may be used as structural probes, but also in the development of rapid assays suitable for large scale screening for PrP isoforms.
Project Documents
• Final Report : Use of novel recombinant prion proteins to generate range of antibodies and peptides that selectively bind aminoterminus   (717k)
Time-Scale and Cost
From: 2001

To: 2004

Cost: £338,467
Contractor / Funded Organisations
University - Reading
Animal Health              
Plants and Animals              
Fields of Study
Animal Health