2011
DOI: 10.1371/journal.ppat.1001277
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Highly Efficient Protein Misfolding Cyclic Amplification

Abstract: Protein misfolding cyclic amplification (PMCA) provides faithful replication of mammalian prions in vitro and has numerous applications in prion research. However, the low efficiency of conversion of PrPC into PrPSc in PMCA limits the applicability of PMCA for many uses including structural studies of infectious prions. It also implies that only a small sub-fraction of PrPC may be available for conversion. Here we show that the yield, rate, and robustness of prion conversion and the sensitivity of prion detect… Show more

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Cited by 92 publications
(108 citation statements)
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“…similar to PMCA conditions). Sonication with beads has been shown to efficiently fragment amyloid fibrils of recombinant PrP into smaller particles (34). We found that the FTIR spectra from "mildly" or "extensively" sonicated samples showed no detectable differences (Fig.…”
Section: Ir-msp Was Not Influenced By the Size Of Prpmentioning
confidence: 82%
See 1 more Smart Citation
“…similar to PMCA conditions). Sonication with beads has been shown to efficiently fragment amyloid fibrils of recombinant PrP into smaller particles (34). We found that the FTIR spectra from "mildly" or "extensively" sonicated samples showed no detectable differences (Fig.…”
Section: Ir-msp Was Not Influenced By the Size Of Prpmentioning
confidence: 82%
“…This discrepancy could be explained by the fact that we used beads for PMCA, which significantly increases the efficiency and robustness of PMCA, as described elsewhere (26,34). In addition, IR-MSP of highly purified protein samples substantially improved the collection and interpretation of IR spectroscopic data as well as the sensitivity of IR analyses.…”
Section: Different Resistance Of 263k-brain 263k-pmca and 263k-pmcamentioning
confidence: 99%
“…Protein Misfolding Cyclic Amplification with Beads-10% NBH from healthy hamsters was prepared as described before (26) and used as a substrate for PMCA with beads (PMCAb) (30). To detect PrP Sc in the brains of inoculated animals, 10 l of 10% BH were added to the 90 l of NBH.…”
Section: Methodsmentioning
confidence: 99%
“…The first modification was automation, which increases the efficiency and sensitivity of conversion (209). The second modification was the addition of Teflon beads, which enhances the conversion to PrP Sc and increases the sensitivity of prion detection (210). The third modification of the PMCA protocol supports the amplification of hamster PrP Sc in the presence of recombinant hamster PrP instead of using brain PrP C as a substrate (211).…”
Section: Protein Misfolding Cyclic Amplification (Pmca)mentioning
confidence: 99%