NR ATJK
AU Kuczius,T.; Grassi,J.; Karch,H.; Groschup,M.H.
TI Enzymatic Degradation of Prion Proteins by the Use of A Composition of Four Common Proteases
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Decontamination DEC-17
PT Konferenz-Poster
AB Prion diseases are neurodegenerative disorders of humans known as Creutzfeldt-Jakob disease (CJD) and its new variant vCJD. The risks of prion transmission via surgical instruments are of current public concern and represent a substantial hazard for the medical system and for the food and farming industry. Security and confidence of patients and consumers has to be improved as well as the very intensive costs of medical instruments used for CJD patients have to be decreased. During pathogenesis the host encoded cellular prion protein (PrPc) is converted into its pathological isoform PrPsc which is the specific marker for the disease and the essential infectious agent. In contrast to PrPc, PrPsc is remarkable resistant to chemicals, heat, proteases and inactivation and decontamination procedures. Thus, there is great demand and urgency for mild decontamination procedures relating to medical and dental instruments as well as to food and medical product safety. In order to develop mild decontamination procedures we have focused our experimental set up to enzymatic treatment using the four common proteases Proteinase K, Papain, Chymotrypsin and Bromelain. As of their various cleavage sites, treatment of PrPsc resulted in specific banding patterns for each single enzyme. Using a composition of all four enzymes in one sample the protein signals dramatically decreased. However, after pre-denaturing by SDS and pre-heating up to 99°, PrPsc of homogenates (10%) derived from human CJD and sheep scrapie were hydrolyzed afterwards by enzyme composition towards levels which are below the detection limit of immunoblot analysis using the sensitive antibodies SAF34, p4, Pri308, 12F10 and SAF70. Even after a ten fold protein concentration by precipitation techniques no protein signals were detectable. Our results emphasize that a combination of denaturing conditions followed by enzymatic treatment is a promising alternative for prion decontamination.
IN Mit einem Gemisch aus vier bekannten Proteasen (Proteinase K, Papain, Chymotrypsin and Bromelain) erzielten die Autoren einen "dramatischen" PrPsc-Abbau, der noch gesteigert wurde, wenn die Hirnhomogenate vor der Enzymbehandlung mit SDS und 99° behandelt wurden. Selbst nach einer zehnfachen Proteinaufkonzentrierung durch Präzipitation lag danach die restliche PrPsc-Konzentration unter der Nachweisgrenze.
AD T.Kuczius, H.Karch, Institute for Hygiene, University Hospital Münster; J.Grassi, Commissariat á l'Energie Atomique (CEA), Service de Pharmacologie et d'Immunologie, CEA/Saclay; M.H.Groschup, Friedrich-Loeffler-Institute, Institute for Novel and Emerging Infectious Diseases
SP englisch
PO Deutschland