Physical Characterization of a Highly Infectious Monodisperse Preparation of TSE Infectivity as a Substrate for Diagnostic Development
Abstract
Purification and characterization of the infectious agent that causes transmissible spongiform encephalopathies would be a breakthrough in the development of diagnostic tests and treatments for the victims of these fatal neurological diseases. Using a new preparation of highly dispersed, nanofiltered scrapie infected brain homogenate, we have determined the buoyant density and sedimentation constant of PrPres, the only known marker for scrapie infectivity. Our sucrose step gradient procedure, which combines the ultracentrifugation approaches of equilibrium density and sedimentation rate, succeeded in both purifying and concentrating both PrPres and infectivity. Scaled up production of purified scrapie infectivity involving one liter of dispersed and nanofiltered scrapie homogenate is underway. We are actively pursuing the original plan of this project, which is to use equilibrium density, sedimentation rate, and other biochemical characteristics to concentrate, purify, and study the scrapie infectious agent. The techniques used are unbiased in their ability to isolate infectious particles, whether they are purely proteinaceous or contain other factors.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2006
- Accession Number
- ADA469533
Entities
People
- Andrew G. Timmes
- Irina Alexeeva
- Luisa L. Gregori
- Robert G. Rohwer