NR AUSQ
AU Löschner,C.; Thomzig,A.; Beekes,M.
TI Amplification of PrPres from hamster-adapted BSE- and Scrapie-strains by Protein Misfolding Cyclic Amplification (PMCA)
QU TSE-Forum, 6. Kongress - Nationale TSE-Forschungsplattform, Greifswald 26.6.-28.6.2006, Poster: Diagnostik DIA-06
PT Konferenz-Poster
AB
Recently, a method for the in vitro amplification of misfolded prion proteins has been published (Soto et al., Nature, 2001) and designated as Protein Misfolding Cyclic Amplification (PMCA). The PMCA-reaction consists of amplification cycles which are composed of two different phases: an incubation phase which allows the growing of PrPres multimers from minute amounts of PrPres in the presence of a large excess of PrPc and a sonication phase, breaking down the PrP-multimers into smaller pieces to multiply the number of converting units.
In order to test whether PrPres derived from different hamster-adapted TSE-strains can be amplified by using PMCA and whether the amplification products show strain-specific characteristics regarding the electrophoretic mobilities and glycosylation patterns of PrP27-30 of the initially introduced material, we used two hamster-adapted scrapie strains (ME7-H and 263K), and a BSE-isolate (BSE-H) as seeds.
In this study, 1x10-7 brain equivalents from terminally ill hamsters infected with 263K, ME7-H or BSE-H were diluted in 10% brain homogenates from healthy hamsters and subsequently subjected to PMCA. Various combinations of temperature, sonication intensity and incubation time were examined for their efficiency in promoting PMCA with these hamster-adapted TSE strains. After proteinase K digestion, the yield and fidelity of amplification was examined by PrPres immunoblotting using mAB 3F4. Results of these studies will be presented.
AD Corina Löschner, Achim Thomzig and Michael Beekes, Robert Koch-Institut, P24, Nordufer 20, 13353 Berlin
SP englisch
PO Deutschland
EA Photo des in Greifswald gezeigten Posters, dessen Titel nach Angabe von Achim Thomzig dem etwas abweichenden Titel im Tagungsband vorzuziehen ist., oben, Details 1, Details 2
OR Tagungsband