NR AJNL
AU Polymenidou,M.; Verghese-Nikolakaki,S.; Groschup,M.H.; Chaplin,M.J.; Stack,M.J.; Plaitakis,A.; Sklaviadis,T.K.
TI A short purification process for quantitative isolation of PrPsc from naturally occurring and experimental transmissible spongiform encephalopathies
QU BMC Infectious Diseases 2002 Oct 8; 2(1): 23
IA http://www.biomedcentral.com/1471-2334/2/23
PT journal article
AB 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. CONCLUSION: 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.
AD Magdalini Polymenidou (Magdalini.Polymenidou@pty.usz.ch), Susan Verghese-Nikolakaki (sverg@pharm.auth.gr), Theodoros Sklaviadis (sklaviad@auth.gr), Laboratory of Pharmacology, Department of Pharmaceutical sciences School of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece; Martin Groschup (mrtin.groschup@rie.bfav.de), Institute of Novel and Emerging Infectious Diseases, Federal Research Centre for Virus Diseases of Animals, Boddenblick 5a17498 Insel Riems, Mecklenburg-Vorpommern, Germany; Melanie J. Chaplin (Melanie.M.J.Chaplin@vla.defra.gsi.gov.uk), Mick J. Stack (m.j.stack@vla.defra.gsi.gov.uk), Veterinary Laboratories Agencies, Weybridge, UK; Andreas Plaitakis (plaitakis@med.uch.gr), Department of Neurology, University of Crete, School of Health Sciences, Heraklion, Crete, Greece
SP englisch