Skip to main content
ARS Home » Research » Publications at this Location » Publication #61284
Title: A COMPETITION IMMUNOASSAY TO DETECT THE SCRAPIE PRION PROTEIN USING FLUORESCENT LABELED PEPTIDES AND CAPILLARY ELECTROPHORESIS.

Author
item Schmerr, Mary Jo
item Cutlip, Randall
item Goodwin, Kathryn
item JENNY, ALLEN - USDA-NVSL-APHIS, AMES, IA

Submitted to: Application High Performance Liquid Chromatography and Capillary Electropho
Publication Type: Abstract Only
Publication Acceptance Date: 9/6/1995
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Scrapie in sheep and in goats belongs to a group of transmissible spongiform encephalopathies that are characterized by neurodegeneration and eventually death. A feature of these diseases is the accumulation in the brain of rod shaped fibrils that form from an aggregated protein. This protein is a protease-resistant form of a normal host cell protein that is found predominantly on neurons. Scrapie infection in sheep and other animals is diagnosed by detection of the prion protein by immunoblot and by immunohistochemistry. In this report, we used capillary electrophoresis and fluorescent labeled peptides to detect the prion protein through competition for a labeled peptide in immunocomplex formation. We used synthesized peptides from positions 89-103 and 142-154 of the prion protein, antiserums that were made to these peptides and extracts from brains of normal and scrapie infected sheep. The parameters that affect immunocomplex formation were studied, by varying the amount of antiserum mixed with the labeled peptide and by using different buffers in the separation. As increasing amounts of unlabeled peptide were added to the assay, a concentration dependent reduction in the immunocomplex peak was observed. The assay detected pg of unlabeled peptide and ng amounts of prion protein in samples from scrapie infected sheep brains. This amount is less than that required for present diagnostic tests.