Detection of bovine spongiform encephalopathy, ovine scrapie prion-related protein (PrPSc) and normal PrPc by monoclonal antibodies raised to copper-refolded prion protein Available to Purchase

Prion-related protein (PrP) is a glycosylphosphatidylinositol-linked cell-surface protein expressed by a wide variety of cells, including those of the nervous system and the immune system. Several functions of normal cellular PrP (PrP^c) have been proposed that may be associated with the capacity of this protein to bind copper. In the present study, we describe the generation of a panel of monoclonal antibodies raised to copper-refolded PrP, which may be used to analyse the normal and disease-associated forms of this protein. The anti-PrP monoclonal antibodies were reactive by Western blot and ELISA with recombinant murine PrP^c refolded in the presence or absence of either copper or manganese, and with the disease-susceptible allelic form V₁₃₆R₁₅₄Q₁₇₁ ('VRQ'; where single-letter amino-acid notation has been used) and disease-resistant allelic form A₁₃₆R₁₅₄R₁₇₁ ('ARR') of recombinant ovine PrP^c. FACS analysis of lymphoid cells using these monoclonal antibodies showed that wild-type non-activated mouse lymphocytes expressed little, if any, PrP^c. These monoclonal antibodies were shown to react with the unglycosylated and monoglycosylated forms of PrP^Sc (abnormal disease-specific conformation of PrP) in prion-infected tissue samples from all of the different species tested by Western blot. In addition, this analysis allowed one to make a distinction between bovine spongiform encephalopathy ('BSE') and scrapie PrP^Sc isolates from experimentally infected sheep on the basis of their different electrophoretic mobilities.