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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #95194
Title: PURIFICATION OF THE PRION PROTEIN FROM SHEEP BRAIN USING HYDROPHILIC INTERACTION CHROMATOGRAPHY

Author
item Schmerr, Mary Jo
item ALPERT, ANDREW - POLY LC,INC.,COLUMBIA, MD
item JENNY, ALLEN - NVSL, PL, AMES, IA

Submitted to: Journal of Chromatography
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/22/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Isolation and testing for the disease agent of scrapie has been difficult with many obstacles and problems. In this research, a new kind of chromatography was developed to extract the disease agent of scrapie. This procedure has made it possible to test for scrapie using tests that are done in ordinary diagnostic laboratories. The procedures in this study ymay result in a practical analytical assay for scrapie in sheep.

Technical Abstract: Scrapie is a prion disease in sheep and goats. Prion diseases are caused by a conformational change of the normal host prion protein to an abnormal form of this protein. The detergents used to extract the protein from tissues interfere with subsequent immunoassays that are used to test for the prion protein. A method was developed to purify the abnormal prion protein using hydrophilic interaction chromatography (HILIC). Tissue samples including brain and lymph nodes were processed with detergent and proteinase K in the standard manner. The resulting extracts were applied to a HILIC column and eluted with a decreasing gradient of acetonitrile in 0.1 percent Trifluoroacetic acid and 50mM hexafluoro-2- propanol. Recovery from the column was approximately 75 percent as determined with radioiodinated prion protein. After drying, the collected peak fractions were resuspended in water and assayed with antibodies specific for the prion protein. The method permitted efficient purification of the prion protein as well as testing by immunoassay, since the interfering detergents were removed.