NR AWKS
AU Moroncini,G.; Mazza,M.; Martucci,F.; Abalos,G.C.; Gazzuola,P.; Corona,C.; Gabrielli,A.; Caramelli,M.; Williamson,R.A.; Casalone,C.
TI Species reactivity of PrP motif-grafted 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-38
PT Konferenz-Poster
AB In order to further assess the species reactivity of PrPsc-specific hybrid human IgG monoclonal antibodies containing the 89-112 or the 136-158 polypeptide motif of mouse prion protein (Moroncini G. et al., Proc Natl Acad Sci USA. 2004 101: 10404-9), we performed immunoprecipitation experiments using brain homogenates prepared from normal and prion-infected (BSE, BASE, Scrapie) bovines and sheep. Binding of IgG 89-112 or IgG 136-158 to PrPc in normal bovine or ovine brain was not detected. Conversely, both motif-grafted reagents immunoprecipitated three PrP bands from undigested and pK-digested BSE, BASE and Scrapie brain extracts. Interestingly, the immunoprecipitated PrP bands, as detected by Western Blot, mirrored the canonical WB profile of the three different prion strains assayed, indicating strain-specific binding of these antibodies. Under the same experimental conditions, the parental human IgG antibody did not show any reactivity with PrPsc nor with PrP 27-30. In conclusion, the data coming from this study, together with those previously published, identify two independent regions of mouse PrP sequence that possess intrinsic specificity and affinity for epitopes found exclusively on PrPsc and PrP 27-30 molecules encountered in prion diseases of animals and humans. Grafting such polypeptides into an IgG scaffold imparted upon this antibody the ability to bind to disease-associated PrP isoforms produced by different prion strains (79A, RML, 263K, BSE, BASE, Scrapie) and in different species. The hybrid antibody reagents we have generated may find broad application in the detection of infectious prions in human and animal materials.
AD G. Moroncini, A. Gabrielli: Sezione di Clinica Medica, Dipartimento di Scienze Mediche e Chirurgiche, Università Politecnica delle Marche, Ancona, Italy; M. Mazza, F. Martucci, P. Gazzuola, C. Corona, M. Caramelli, C. Casalone: CEA, Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Torino, Italy; G. C. Abalos, R. A. Williamson: Department of Immunology, The Scripps Research Institute, La Jolla, CA, USA. E-mail: g.moroncini@univpm.it
SP englisch
PO Italien