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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #90012
Title: PURIFICATION OF THE PRION PROTEIN FROM SHEEP BRAIN USING HYDROPHILIC INTERACTION CHROMATOGRAPHY

Author
item Schmerr, Mary Jo
item ALPERT, ANDREW - POLYLC,INC.,COLUMBIA, MD.
item JENNY, ALLEN - NVSL, APHIS, AMES, IA

Submitted to: Association of Biomolecular Resources Facilities Symposium
Publication Type: Abstract Only
Publication Acceptance Date: 3/25/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary:

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 immunosassays 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 TFA 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.