title: A short purification process for quantitative isolation of PrPSc from naturally occurring and experimental transmissible spongiform encephalopathies.
abstract: BACKGROUND
  Transmissible spongiform encephalopathies (TSEs) are neurodegenerative diseases affecting both humans and animals. They are associated with post-translational conversion of the normal cellular prion protein (PrPC) into a heat- and protease-resistant abnormal isoform (PrPSc). Detection of PrPSc in individuals is widely utilized for the diagnosis of prion diseases.
  
  
  METHODS
  TSE brain tissue samples have been processed in order to quantitatively isolate PrPSc. The protocol includes an initial homogenization, digestion with proteinase K and salt precipitation.
  
  
  RESULTS
  Here we show that over 97 percent of the PrPSc present can be precipitated from infected brain material using this simple salting-out procedure for proteins. No chemically harsh conditions are used during the process in order to conserve the native quality of the isolated protein.
  
  
  CONCLUSIONS
  The resulting PrPSc-enriched preparation should provide a suitable substrate for analyzing the structure of the prion agent and for scavenging for other molecules with which it may associate. In comparison with most methods that exist today, the one described in this study is rapid, cost-effective and does not demand expensive laboratory equipment.
authors.affiliation: Laboratory of Pharmacology, Department of Pharmaceutical sciences School of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece. magdalini.polymenidou@pty.usz.ch
authors.forename: Magdalini
authors.initials: M
authors.lastname: Polymenidou
authors.forename: Susan
authors.initials: S
authors.lastname: Verghese-Nikolakaki
authors.forename: Martin
authors.initials: M
authors.lastname: Groschup
authors.forename: Melanie J
authors.initials: MJ
authors.lastname: Chaplin
authors.forename: Mick J
authors.initials: MJ
authors.lastname: Stack
authors.forename: Andreas
authors.initials: A
authors.lastname: Plaitakis
authors.forename: Theodoros
authors.initials: T
authors.lastname: Sklaviadis
mesh_terms: D000818:Animals; D011232:Chemical Precipitation; D004195:Disease Models, Animal; D006801:Humans; D051379:Mice; D018620:PrPSc Proteins / Q000032:analysis / Q000302:isolation & purification*; D017096:Prion Diseases / Q000378:metabolism*; D016622:Silver Staining; D013045:Species Specificity
chemical_list: D018620:PrPSc Proteins