Establishment of bovine prion peptide-based monoclonal antibodies for identifying bovine prion

Abstract

To obtain high titer monoclonal antibodies (McAbs) which can react with mammalian prion protein (PrP), Balb/C mice were immunized with bovine (Bo) PrP peptide (BoPrP 209-228 aa) coupled to keyhole limpet hemocyanin (KLH). The hybridoma cell lines secreting monoclonal antibodies against the peptide were established by cell fusion and cloning. The obtained McAbs were applied to detect recombinant human, bovine and hamster PrP, cellular prion protein (PrP(c)) in normal bovine brain and pathogenic scrapie prion protein (PrP(Sc)) accumulated in the medulla oblongata of bovine spongiform encephalopathy(BSE)specimen with Western blot and immunohistochemical detection, respectively. The current procedure might offer a simple, feasible method to raise high titer antibodies for studying biological features of PrP in mammals, as well as detection of transmissible spongiform encephalopathy (TSE) and diagnosis of BSE, in particular.

Conventional and State-of-the-Art Detection Methods of Bovine Spongiform Encephalopathy (BSE)

“…The reactivity of mAb to prion protein also depends on the analytical method, as different antigen treatments in different methods resulted in the exposition of certain epitopes in a conformation-dependent manner and affected the affinity of antibodies and antigens. For example, the B4 antibody reacted with both PrP C and PrP Sc in the IHC test, but it showed a faint reaction with PrP C in the Western blot test [ 42 ].…”

Progress in the development of therapeutic antibodies targeting prion proteins and β-amyloid peptides

“…Several anti-PrP [33] and anti-Aβ [34] antibodies have been developed by our group, and some of these may be useful for diagnosis and treatment of AD. The A8 MAb that preferentially recognizes Aβ oligomers of about 16-25 kD can neutralize the cytotoxicity induced by Aβ oligomers, thus inhibiting fiber maturity and improving the memory of the AD mouse model.…”