Proteolytic inactivation of the bovine spongiform encephalopathy agent
| Autor: | Clive M. Buswell, Jean Carr, Neil D.H. Raven, Graham Hall, J. Mark Sutton, Anne H. McLeod, Jo Dickinson, David M. Taylor, Mike Dennis, Heather Murdoch |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Proteases
Prions animal diseases medicine.medical_treatment Blotting Western Biophysics Biochemistry Microbiology Incubation period Mice Western blot Endopeptidases Enzyme Stability medicine Bioassay Animals Molecular Biology Incubation Decontamination Protease Transmissible spongiform encephalopathy biology medicine.diagnostic_test Temperature Brain Cell Biology Hydrogen-Ion Concentration medicine.disease Survival Analysis nervous system diseases Encephalopathy Bovine Spongiform biology.protein Biological Assay Cattle Antibody |
| Zdroj: | Biochemical and biophysical research communications. 317(4) |
| ISSN: | 0006-291X |
| Popis: | Thermostable proteases have been investigated for their ability to provide a novel biological solution to decontamination of prion agents responsible for transmissible spongiform encephalopathies (TSEs). Proteases were identified that digested total mouse brain homogenate (MBH) protein from uninfected mice. These proteases were then evaluated for digestion of BSE (301V) infectious MBH over a range of pH and temperatures, screened for loss of anti-prion antibody 6H4 immunoreactivity and protease-treated infectious MBH assessed in mouse bioassay using VM mice. Despite a number of proteases eliminating all 6H4-immunoreactive material, only the subtilisin-enzyme Properase showed a significant extension in incubation period in mouse bioassays following a 30-min incubation at 60 degrees C and pH 12. These results demonstrate the potential of the method to provide a practical solution to the problems of TSE contamination of surgical instruments and highlight the inadequacy of using Western blot for assessment of decontamination/inactivation of TSE agents. |
| Databáze: | OpenAIRE |
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