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• W2041529800 abstract "Highly sensitive in vitro screening tests are required to prevent the iatrogenic spread of variant Creutzfeldt–Jakob disease (vCJD). Protein misfolding cyclic amplification (PMCA) is a candidate for such a test, but the sensitivity of this method is insufficient. Polyanions were reported to enhance PMCA efficiency, but their effects on vCJD are unclear. We developed a cell-PMCA of vCJD, wherein cell lysate containing exogenously expressed human PrP was used as substrates, to investigate the effects of various sulfated polysaccharides on amplification efficiency. PrPres amounts after cell-PMCA were analyzed by western blotting. Heparin, dermatan sulfate, and dextran sulfate (average molecular weight [MW] 1400 kDa) enhanced efficiency, but dextran sulfate (average MW 8 kDa) and a heparin pentasaccharide analog had no effect. Pentosan polysulfate inhibited cell-PMCA reaction. The amplification efficiency of cell-PMCA of vCJD increased to >100-fold per round with heparin. The enhancing effects of heparin on cell-PMCA were seed dependent: it was high for vCJD, low for sporadic Creutzfeldt–Jakob disease, and low to negligible for hamster-adapted scrapie-derived 263 K. In multi-round PMCA, signals were detected at earlier rounds with heparin than without heparin, and PrPSc in 10−10 diluted vCJD brain was detected by the sixth round. Heparin-assisted cell-PMCA of vCJD represents a significant step toward detecting very minute amounts of PrPSc in the body fluids of asymptomatic vCJD patients." @default.
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• W2041529800 date "2011-07-01" @default.
• W2041529800 modified "2023-10-10" @default.
• W2041529800 title "Heparin enhances the cell-protein misfolding cyclic amplification efficiency of variant Creutzfeldt–Jakob disease" @default.
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• W2041529800 doi "https://doi.org/10.1016/j.neulet.2011.04.072" @default.
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