Highly sensitive, quantitative cell-based assay for prions adsorbed to solid surfaces

Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3479-83. doi: 10.1073/pnas.0813342106. Epub 2009 Feb 9.

Abstract

Prions are comprised principally of aggregates of a misfolded host protein and cause fatal transmissible neurodegenerative disorders of humans and animals, such as variant Creutzfeldt-Jakob disease and bovine spongiform encephalopathy. Prions pose significant public health concerns, including contamination of blood products and surgical instruments; require laborious and often insensitive animal bioassay to detect; and resist conventional hospital sterilization methods. A major experimental advance was the cell culture-based scrapie cell assay, allowing prion titres to be estimated more precisely and an order of magnitude faster than by animal bioassays. Here we describe a bioassay method that exploits the marked binding affinity of prions to steel surfaces. Using steel wires as a concentrating and sensitization tool and combining with an adapted scrapie cell endpoint assay we can achieve, for mouse prions, a sensitivity 100x higher than that achieved in standard mouse bioassays. The rapidity and sensitivity of this assay offers a major advance over small animal bioassay in many aspects of prion research. In addition, its specific application in assay of metal-bound prions allows evaluation of novel prion decontamination methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Prions / analysis*
  • Prions / metabolism*
  • Protein Stability
  • Substrate Specificity
  • Temperature

Substances

  • Prions