Preparation and characterization of antibodies against mouse prion protein (PrP) peptides.
Antisera were raised in rabbits against three peptides, representing amino acid sequences 150 to 159, 165 to 174, and 213 to 226 of mouse prion (PrP), which were synthesized by using a multiple antigenic peptide (MAP) system. The reactivities of these sera to PrP were examined by an enzyme-linked immunosorbent assay (ELISA), Western immunoblotting (WB), and immunohistochemical procedures. The results of both ELISA and WB showed that antisera to peptide sequence 150 to 159 (Ab150-159) did not react with purified mouse PrP. On the other hand, sera to the sequence 165 to 174 (Ab165-174) reacted weakly with purified mouse PrP, as detected by WB but not by ELISA. However, antiserum to peptide sequence 213 to 226 (Ab213-226) reacted strongly with mouse, Syrian hamster, and sheep PrP by WB and with mouse PrP as shown by the results of ELISA. Moreover, Ab213-226 clearly detected PrP immunohistochemically in mouse, Syrian hamster, and sheep brains affected with scrapie as well as in the brain of a cow with bovine spongiform encephalopathy. From these data, we conclude that rabbit antiserum against the MAP representing amino acid sequence 213 to 226 of mouse PrP is useful as a diagnostic tool for prion disease of animals.
Yokoyama
T
T
National Institute of Animal Health, Ibaraki, Japan.
Kimura
K
K
Tagawa
Y
Y
Yuasa
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D001923:Brain Chemistry; D002417:Cattle; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D005260:Female; D006168:Guinea Pigs; D007106:Immune Sera; D007150:Immunohistochemistry; D008647:Mesocricetus; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000138:chemical synthesis* / Q000276:immunology*; D011328:Prions / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D011487:Protein Conformation; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D013045:Species Specificity
D000906:Antibodies; D000939:Epitopes; D007106:Immune Sera; D010446:Peptide Fragments; D011328:Prions