Abstract
BackgroundTransmissible 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.MethodsTSE 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.ResultsHere 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.ConclusionThe 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.
Highlights
Transmissible spongiform encephalopathies (TSEs) are neurodegenerative diseases affecting both humans and animals
The disease is characterized by long incubation periods which, as in the recent crossover of the bovine spongiform encephalopathy (BSE) agent to humans, may be prolonged further when the disease is transmitted from one species to another
BSE brain samples were obtained from VLA, Weybridge, UK and the human Creutzfeldt-Jakob disease (CJD) brain samples came from recently reported cases of sporadic CJD from within Greece [13]
Summary
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). Bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease (CJD) are all related transmissible spongiform encephalopathies, the common major causative agent of which is believed to be a protease- and heat-resistant, beta-sheet rich isoform (PrPSc) of the normal cellular prion protein (PrPC) [1]. The disease is characterized by long incubation periods which, as in the recent crossover of the bovine spongiform encephalopathy (BSE) agent to humans, may be prolonged further when the disease is transmitted from one species to another. The ultimate confirmation of the disease comes only after post-mortem examination of the brain, even though extensive research being carried out in the field offers hope for pre-clinical diagnosis through the detection of PrPC in tonsils or body fluids
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