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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #99734
Title: THE USE OF CAPILLARY ELECTROPHORESIS AND FLUORESCENT LABELED PEPTIDES TO DETECT THE ABNORMAL PRION PROTEIN IN THE BLOOD OF ANIMALS THAT ARE INFECTED WITH A TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY

Author
item Schmerr, Mary Jo
item JENNY, ALLEN - NVSL, PL., AMES, IA.
item BULGIN, MARIE - UNIV.OF IDAHO,CALDWELL,ID
item Miller, Janice
item Hamir, Amirali
item Cutlip, Randall
item Goodwin, Kathryn

Submitted to: Journal of Chromatography
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/1/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: A sensitive diagnostic test is needed to identify animals that are infected with a transmissible spongiform encephalopathy. A sensitive assay was developed that uses capillary electrophoresis. This assay can detect the agent that causes transmissible spongiform encephalopathies in the blood of infected animals. This research may lead to a diagnostic test that will lead to methods to control and eventually eradicate these diseases. This will prevent economic losses to the animal producers and industries such as the pharmaceutical and cosmetic industries that use animal products and protect the human and animal food supply.

Technical Abstract: Transmissible spongiform encephalopathies in humans and in animals are fatal neuro-degenerative diseases with long incubation times. The putative cause of these diseases is a normal host protein, the prion protein, that becomes altered. This abnormal prion protein is found mostly in the brains of infected individuals in later stages of the disease, but also can be found in lymphoid and other tissues in lower amounts. In order to eradicate this disease in animals, it is important to develop a system that can concentrate the abnormal prion protein and an assay that is very sensitive. A peptide from the carboxyl terminal region, amino acid positions 218-232, was labeled with fluorescein during the synthesis of the peptide at the amino terminus. Antibodies that have been produced to this peptide were affinity purified and used in a capillary electrophoresis immunoassay. The amount of fluorescein labeled peptide in the capillary was 50 attomoles. Blood was obtained from normal sheep and elk and from sheep infected with scrapie and elk infected with chronic wasting disease. Buffy coats and plasma were prepared by a conventional method. After treatment with proteinase K, which destroys the normal protein but not the altered one, the blood fractions were extracted and tested in the capillary electrophoresis immunoassay for the abnormal prion protein. The abnormal prion protein was detected in fractions from blood from infected animals but not from normal animals. This assay makes a pre-clinical assay possible for these diseases and might be adapted to test for the abnormal prion protein in process materials that are used for manufacture of pharmaceuticals and products for human and animal consumption.