Physical, chemical and kinetic factors affecting prion infectivity. Issue 3 (3rd May 2016)

• Record Type: Journal Article
• Authors: Properzi, Francesca; Badhan, Anjna; Klier, Steffi; Schmidt, Christian; Klöhn, Peter C.; Wadsworth, Jonathan D. F.; Clarke, Anthony R.; Jackson, Graham S.; Collinge, John
• Is Part Of: Prion. Volume 10:Issue 3(2016)
• Page Start: 251
• Page End: 261
• Publication Date: 2016-05-03
• Subjects: BSE -- CJD -- cell culture -- prion -- PrP -- RML -- scrapie; Protein folding -- Periodicals; Prions -- Periodicals; Proteins -- Biotechnology -- Periodicals; 572.633
• DOI: 10.1080/19336896.2016.1181250
• Languages: English
• ISSNs: 1933-6896

Abstract

ABSTRACT: The mouse-adapted scrapie prion strain RML is one of the most widely used in prion research. The introduction of a cell culture-based assay of RML prions, the scrapie cell assay (SCA) allows more rapid and precise prion titration. A semi-automated version of this assay (ASCA) was applied to explore a range of conditions that might influence the infectivity and properties of RML prions. These include resistance to freeze-thaw procedures; stability to endogenous proteases in brain homogenate despite prolonged exposure to varying temperatures; distribution of infective material between pellet and supernatant after centrifugation, the effect of reducing agents and the influence of detergent additives on the efficiency of infection. Apparent infectivity is increased significantly by interaction with cationic detergents. Importantly, we have also elucidated the relationship between the duration of exposure of cells to RML prions and the transmission of infection. We established that the infection process following contact of cells with RML prions is rapid and followed an exponential time course, implying a single rate-limiting process.