|Schmerr, Mary Jo|
|Jenny, Allen - APHIS NVSL PL AMES IA|
Submitted to: Proceedings of the International Symposium on Capillary Electrophoresis
Publication Type: Abstract Only
Publication Acceptance Date: November 18, 1997
Publication Date: N/A
Technical Abstract: Prion diseases or transmissible spongiform encephalopathies belong to a group of neurodegenerative diseases that infect both animals and humans. These diseases are associated with the accumulation of fibrils in the brains of affected individuals. These fibrils are composed of an abnormal isoform of a host-encoded glycoprotein that is characterized by its insolubility and partial resistance to proteases. A post translational mechanism which involves a conformational change from about 40 per cent alpha-helix and about 3 per cent beta-pleated sheet in the normal form to 40 per cent beta-pleated sheet and 30 per cent alpha-helix in the altered form occurs. When specific antisera to the prion protein is used to make an immunoblot of this altered protein, three characteristic isoforms are observed. The migration of these isoforms has been suggested as a means to determine the strain of the prion protein. We explored the use of capillary yisoelectrofocusing to differentiate between scrapie prion protein from sheep and scrapie prion protein that has been adapted to hamsters. We used a Beckman 5500 P/ACE using UV detection at 280nm. A cIEF 3-10 Kit from Beckman Instruments was used to perform the analysis. The scrapie prion protein was solubilized in 95 per cent isopropyl alcohol containing 20 mM ammonium formate pH 3.5 at 56degrees C. The solubilized scrapie prion protein was placed over a high performance hydrophilic interaction column. After elution, the peaks were assayed for immunoreactivity with specific antisera. The peaks that contained immunoreactivity were then placed on the cIEF capillary. The pIs of the samples varied depending upon the source of the scrapie prion protein. The pIs ranged from 6.0 to 4.5.