NR AQLI
AU McLeod,A.H.; Murdoch,H.; Dickinson,J.; Dennis,M.J.; Hall,G.A.; Buswell,C.M.; Carr,J.; Taylor,D.M.; Sutton,J.M.; Raven,N.D.H.
TI Proteolytic inactivation of the bovine spongiform encephalopathy agent
QU Biochemical and Biophysical Research Communications 2004 May 14; 317(4): 1165-70
PT journal article
AB 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 ° 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.
MH Animals; Biological Assay; Blotting, Western; Brain/metabolism; Cattle; Decontamination; Encephalopathy, Bovine Spongiform/*metabolism; Endopeptidases/*metabolism; Enzyme Stability; Hydrogen-Ion Concentration; Mice; Prions/immunology/*metabolism; Support, Non-U.S. Gov't; Survival Analysis; Temperature
AD Anne H. McLeod, Heather Murdoch, Jo Dickinson, Mike J. Dennis, Graham A. Hall, Clive M. Buswell, Jean Carr, J. Mark Sutton (mark.sutton@hpa.org.uk), Neil D.H. Raven, Health Protection Agency, Porton Down, Salisbury, Wiltshire SP4 0JG, UK; David M. Taylor, Sedecon 2000, 147, Oxgangs Road North, Edinburgh EH13 9DX, UK
SP englisch
PO USA