Comparison of three monoclonal antibodies for use in immunohistochemical detection of bovine spongiform encephalopathy protease-resistant prion protein.
Confirmatory diagnosis of prion diseases in humans and animals relies on the histopathological examination and immunodetection of the protease-resistant isoform of prion protein (PrPres). The generation of novel PrP-specific monoclonal antibodies (MAbs) has greatly improved diagnostic methodology and basic research on prion diseases as well. In this study, the performance of 3 different PrP-specific MAbs in recognizing brain PrPres deposits from cows affected with bovine spongiform encephalopathy (BSE) was compared by using a standard immunohistochemical technique under different pretreatment conditions. All antibodies showed similar reactivity after denaturing treatment. However, greater differences were found among them after proteinase K treatment, even in the absence of a denaturing step. In fact, 1 MAb (2A11) was able to react with PrPres deposits in the absence of a denaturing step, yielding the strongest signal and confirming the usefulness of MAb 2A11 in immunohistochemistry for the diagnosis of BSE.
Salguero
Francisco J
FJ
Centro de Investigación en Sanidad Animal (CISA-INIA), Madrid, Spain. salguero@inia.es
Díaz-San
Segundo Fayna
SF
Brun
Alejandro
A
Cano
María J
MJ
Torres
Juan M
JM
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001933:Brain Stem / Q000737:chemistry / Q000473:pathology; D002417:Cattle; D016643:Encephalopathy, Bovine Spongiform / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D019286:Endopeptidase K / Q000378:metabolism; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D007150:Immunohistochemistry / Q000379:methods / Q000662:veterinary*; D008969:Molecular Sequence Data; D011328:Prions / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D011489:Protein Denaturation
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011328:Prions; D019286:Endopeptidase K