NR AWBK
AU Barbieri,I.; Brocchi,E.; Capucci,L.
TI Further step in the characterization of a new group of monoclonal antibodies
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions DIA-04
PT Konferenz-Poster
AB In the past few years some new monoclonal antibodies have been raised against both recombinant bovine PrP and the synthetic peptide corresponding to the amino acid sequence of bovine PrP 153-172. They have been characterized by western blot and IHC for their ability to recognize cellular and pathological PrP from different animal species (Barbieri et al., Prion 2003, Munich 8-10 October 2003). Here we report on the use of such AcM in the detection of PrP from a set of brain samples from wild ruminants including deer, roe-deer, chamois, fallow-deer and ibex. The AcM directed to the N-terminal part (3G5, 3H1, 4D7), the core region (2B10 and 4E3) and the C-terminal part (4G11) of PrP were able to recognize PrPc from all the samples. Among AcM raised against the synthetic peptide, 3E2 recognized PrP from all samples while 3G6 was able to bind no species. These data are consistent with previous results indicating that 3G6 is specific for bovine/human PrP, while 3E2 has higher affinity for sheep and goat PrP with which wild ruminant PrP share a very high degree of amino acid sequence homology (97.2% to 99.6%). Data on the characterization of a new AcM (4E12) able to specifically recognize the mono and unglycosylated form of both cellular and pathological PrP from different animal species including bovine, sheep, goat, hamster and human will also be presented. AcM were also tested on BASE samples. AcM to the N-terminal part of PrP were unable to detect both BSE and BASE suggesting their epitope is lost after digestion with Pk both in BASE and BSE as expected due to the amino acidic sequence similarity between BSE and BASE PrP (Casalone et al., PNAS, 101, 3065-3070, 2004). AcM to the core of PrP and to C-terminal are able to recognize BASE and the molecular features that distinguish it from BSE such as different glycosylation ratios and lower molecular weight of unglycosylated PrP.
AD Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Brescia, Italy. E-mail: ibarbieri@bs.izs.it
SP englisch
PO Italien