P54 Production and Characterisation of Monoclonal Antibodies Against Human Platelet Prion Protein

Abstract

To date all of the existing monoclonal antibodies which react with human prion protein have been produced following immunisation with recombinant PrP, PrP from other species or synthetic peptide fragments. The aim of this project was to produce monoclonal antibodies raised against the native form of PrPc purified from human platelets. Following immunization of PrP‐/‐ Edinburgh mice and the use of conventional hybridoma techniques a panel of 12 monoclonal antibodies has been established. These antibodies have been characterized by epitope mapping; binding to both native/denatured platelet PrP and native/denatured α‐helical/β‐sheet recombinant mouse PrP; immunoprecipitation of PrPc, PrPsc and PrPres from neurological control/vCJD brain homogenates and as probes for Western blotting, immunohistochemistry and flow‐cytometry studies. A range of epitopes was recognised by the panel, with discrimination between normal α‐helical and abnormal β‐sheet forms. Several of the antibodies specifically captured abnormal forms of prion protein (PrPsc and PrPres ) found in vCJD‐infected CNS and lymphoreticular tissues. It is hoped that these antibodies will aid in the early diagnosis and classification of human prion disease and the detection of vCJD infectivity in tissues for transplant and blood donations.