Decontamination of prion protein (BSE301V) using a genetically engineered protease.
A previous study has demonstrated the potential of alkaline proteases to inactivate bovine spongiform encephalopathy (BSE301V). Here we explored the use of MC3, a genetically engineered variant of Bacillus lentus subtilisin. MC3 was used to digest BSE301V infectious mouse brain homogenate (iMBH). MC3 eliminated all detectable 6H4-immunoreactive material at pH 10 and 12; however, Proteinase K was only partially effective at pH 12. When bioassayed in VM mice, MC3- and Proteinase K-digested iMBH gave respectively 66.6% and 22.7% survival rates. Using a titration series for disease incubation, this equates to a >7log reduction in infectivity for MC3 and >6log reduction for Proteinase K. This study demonstrates the potential for thermostable proteases to be developed as effective inactivation processes for prion agents in healthcare management.
Dickinson
J
J
Health Protection Agency, Porton Down, Salisbury, UK.
Murdoch
H
H
Dennis
M J
MJ
Hall
G A
GA
Bott
R
R
Crabb
W D
WD
Penet
C
C
Sutton
J M
JM
Raven
N D H
ND
D000818:Animals; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D002417:Cattle; D003666:Decontamination / Q000379:methods*; D016643:Encephalopathy, Bovine Spongiform / Q000517:prevention & control; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D051379:Mice; D011328:Prions / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020860:Subtilisin / Q000235:genetics / Q000378:metabolism*; D016019:Survival Analysis
D011328:Prions; D011994:Recombinant Proteins; D020860:Subtilisin