Generation of monoclonal antibody that distinguishes PrPSc from PrPC and neutralizes prion infectivity.
To establish PrP(Sc)-specific mAbs, we immunized Prnp(-/-) mice with PrP(Sc) purified from prion-infected mice. Using this approach, we obtained mAb 6H10, which reacted with PrP(Sc) treated with proteinase K, but not with PrP(Sc) pretreated with more than 3 M GdnHCl. In contrast, reactivity of pan-PrP mAbs increased with increasing concentrations of GdnHCl used for pretreatment of PrP(Sc). In histoblot analysis, mAb 6H10 showed a positive reaction on a non-denatured histoblot but reactivity was lower when the histoblot was pretreated by autoclaving. Epitope analysis suggested that the extreme C-terminus of PrP is likely to be part of the epitope for mAb 6H10. MAb 6H10 immunoprecipitated PrP(Sc) from brains of mice, sheep, and cattle infected with prions. Furthermore, pretreatment of purified PrP(Sc) with mAb 6H10 reduced the infectious titer more than 1 log. Taken together, these results suggest that mAb 6H10 recognizes a conformational epitope on PrP(Sc) that is related to prion infectivity.
Horiuchi
Motohiro
M
Laboratory of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18, Nishi-9, Kita-ku, Sapporo, Hokkadio 060-0818, Japan. horiuchi@vetmed.hokudai.ac.jp
Karino
Ayako
A
Furuoka
Hidefumi
H
Ishiguro
Naotaka
N
Kimura
Kumiko
K
Shinagawa
Morikazu
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D057134:Antibodies, Neutralizing / Q000096:biosynthesis*; D000918:Antibody Specificity; D001483:Base Sequence; D001921:Brain / Q000276:immunology; D001923:Brain Chemistry; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D018621:PrPC Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018620:PrPSc Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017096:Prion Diseases / Q000276:immunology / Q000378:metabolism; D011328:Prions / Q000737:chemistry / Q000276:immunology / Q000472:pathogenicity; D011487:Protein Conformation; D012756:Sheep
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D017931:DNA Primers; D000939:Epitopes; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions