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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #79448

Title: CAPILLARY ELECTROPHORESIS ASSAYS FOR THE SCRAPIE PRION PROTEIN

Author
item Schmerr, Mary Jo
item Cutlip, Randall

Submitted to: National Management Health Care Conference
Publication Type: Proceedings
Publication Acceptance Date: 2/4/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Scrapie in sheep and in goats is the prototype of a group of transmissible spongiform encephalopathies (TSE) that are characterized by neuro- degeneration and eventually death. A feature of these diseases is the accumulation in the brain of rod shaped fibrils that form from an aggregated protein. When the aggregated protein is denatured in SDS and 2-mercaptoethanol, a molecular mass of approximately 25Kdaltons and two minor isoforms are observed. The present analytical test that is used to determine the presence of the prion protein is immunoblot. This is a postmortem test. In order to achieve sensitivities that may be required for an antemortem test, we investigated the possibility of using capillary electrophoresis. This methodology allows for several approaches. We developed a competition immunoassay using fluorescent labeled peptides from the prion protein with laser induced fluorescence for detection. The prion protein was prepared through ultracentrifugation and denaturing with SDS and 2-mercaptoethanol. Removal of the denaturing agents is required before performing this assay. Although this test is at least a 1000 times more sensitive than the other tests, it is limited by the preparation of the prion protein. Other techniques using SDS-gel capillary electrophoresis were explored showing that the scrapie infected brain samples could be differentiated from the normal brain samples.