Chicken antibody against a restrictive epitope of prion protein distinguishes normal and abnormal prion proteins.
Recently, we reported the application of a recombinant chicken IgY monoclonal antibody, Ab3-15, against mammalian prion protein (PrP), for the diagnosis of bovine spongiform encephalopathy in cattle. In this study, we have characterized a soluble, single-chain variable fragment (scFv) form of this antibody, sphAb3-15 using brain homogenates from mice. This sphAb3-15 antibody recognized denatured forms of both PrP(C) and PrP(Sc), and PrP(Sc) after PK-treatment, on Western blotting. In sandwich ELISAs, on dot blots and by immunoprecipitation, sphAb3-15 efficiently bound to PrP from normal brain homogenates, but weakly bound PrP from scrapie-infected brain homogenates. These results suggest that sphAb3-15 selectively recognizes PrP(C) under native conditions and that the epitope recognized by sphAb3-15 may undergo conformational changes during the conversion of PrP(C) into PrP(Sc).
Miyamoto
Kazuyoshi
K
Laboratory of Immunobiology, Department of Molecular and Applied Bioscience, Graduate School of Biosphere Science, Hiroshima University, 1-4-4 Kagamiyama, Higashi-Hiroshima, Hiroshima, Japan.
Kimura
Sota
S
Nakamura
Naoto
N
Yokoyama
Takashi
T
Horiuchi
Hiroyuki
H
Furusawa
Shuichi
S
Matsuda
Haruo
H
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D002417:Cattle; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D051379:Mice; D018621:PrPC Proteins / Q000276:immunology; D018620:PrPSc Proteins / Q000276:immunology; D017096:Prion Diseases / Q000175:diagnosis / Q000276:immunology; D011328:Prions / Q000276:immunology*; D012608:Scrapie / Q000175:diagnosis / Q000276:immunology
D000939:Epitopes; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions