title: Proteolytic inactivation of the bovine spongiform encephalopathy agent.
abstract: 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.
authors.affiliation: Health Protection Agency, Porton Down, Salisbury, Wiltshire SP4 0JG, UK.
authors.forename: Anne H
authors.initials: AH
authors.lastname: McLeod
authors.forename: Heather
authors.initials: H
authors.lastname: Murdoch
authors.forename: Jo
authors.initials: J
authors.lastname: Dickinson
authors.forename: Mike J
authors.initials: MJ
authors.lastname: Dennis
authors.forename: Graham A
authors.initials: GA
authors.lastname: Hall
authors.forename: Clive M
authors.initials: CM
authors.lastname: Buswell
authors.forename: Jean
authors.initials: J
authors.lastname: Carr
authors.forename: David M
authors.initials: DM
authors.lastname: Taylor
authors.forename: J Mark
authors.initials: JM
authors.lastname: Sutton
authors.forename: Neil D H
authors.initials: ND
authors.lastname: Raven
mesh_terms: D000818:Animals; D001681:Biological Assay; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002417:Cattle; D003666:Decontamination; D016643:Encephalopathy, Bovine Spongiform / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D051379:Mice; D011328:Prions / Q000276:immunology / Q000378:metabolism*; D016019:Survival Analysis; D013696:Temperature
chemical_list: D011328:Prions; D010450:Endopeptidases