Generation of Antibodies against Bovine Recombinant Prion Protein in Various Strains of Mice

Abstract

Sc from sheep brain in Western blot assays. The epitope specificity of these MAbs was determined, and applicability to immunohistochemical detection of prions was studied. The MAbs generated will be useful tools in the development of TSE immunochemical diagnosis and for research. This is the first report of the development of anti-PrP MAbs by use of autoimmune NZB/NZW F 1 mice as an alternative approach for the generation of PrP-specific MAbs.

Identification of an epitope on the recombinant bovine PrP that is able to elicit a prominent immune response in wild-type mice

“…Our results show that it is actually possible to obtain high anti-PrP antibody titres in healthy mice expressing normal PrP to chemically unmodified bovine PrP not linked to any immunogenic carrier protein in contrast to the recent report from Andrievskaia et al [35], where only the immunization of autoimmunity-prone mice with recBoPrP conjugated to keyhole limpet hemocyanin gave titres comparable to ours. It was shown that the structure of recombinant protein that we used for the immunization is essentially identical to the polypeptide structure of bovine PrP C isolated from the brain [36].…”

Generation of potent mouse monoclonal antibodies to self-proteins using T-cell epitope “tags”

“…SLE-like mouse models (e.g., the NZB/W mouse strain) that display defective B cell tolerance have been successfully employed to generate antibodies to a number of self-antigens and closely-related proteins, but have not been universally successful. 11,12 Alternatively, mice with a genetic knockout for a particular protein can be immunized with that protein exogenously to elicit an immune response. This approach has been used to develop antibodies to mouse and human butyrylcholinesterase 13 and to mouse cellular prion protein.…”

Binding of bovine T194A PrPCby PrPSc-specific antibodies

“…We also used wild-type bovine prion containing amino acid residues (25−242) expressed and purified as described previously. 49 Monoclonal antibody (mAb) M2188 (IgG2b) which recognizes amino acids 146 to 153 (SRPLIHFG) in PrP C was generated as described in Andrievskaia et al 49 Polyclonal antibody (pAb) SN6b, was raised against a highly immunogenic presentation of the following epitope QVYYRPVDQYSNQN in the form (forwardback-back) 4 in sheep and affinity purified (Covance) from sheep serum. 20 Polyclonal antibodies were selected to mimic the situation of vaccination and detect binding to all conformations explored by the mutant T194A of PrP C .…”