Concentration of disease-associated prion protein with silicon dioxide.
Reagents that can precipitate the disease-associated prion protein (PrP(Sc)) are vital for the development of high sensitivity tests to detect low levels of this disease marker in biological material. Here, a range of minerals are shown to precipitate both ovine cellular prion protein (PrP(C)) and ovine scrapie PrP(Sc). The precipitation of prion protein with silicon dioxide is unaffected by PrP(Sc) strain or host species and the method can be used to precipitate bovine BSE. This method can reliably concentrate protease-resistant ovine PrP(Sc) (PrP(res)) derived from 1.69 microg of brain protein from a clinically infected animal diluted into either 50 ml of buffer or 15 ml of plasma. The introduction of a SiO(2) precipitation step into the immunological detection of PrP(res) increased detection sensitivity by over 1,500-fold. Minerals such as SiO(2) are readily available, low cost reagents with generic application to the concentration of diseases-associated prion proteins.
Rees
Helen C
HC
Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, UK.
Maddison
Ben C
BC
Owen
Jonathan P
JP
Whitelam
Garry C
GC
Gough
Kevin C
KC
D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry; D002417:Cattle; D011232:Chemical Precipitation; D006863:Hydrogen-Ion Concentration; D008903:Minerals / Q000737:chemistry; D018621:PrPC Proteins / Q000032:analysis* / Q000097:blood; D018620:PrPSc Proteins / Q000032:analysis* / Q000097:blood; D017096:Prion Diseases / Q000378:metabolism*; D012680:Sensitivity and Specificity; D012756:Sheep; D012822:Silicon Dioxide / Q000737:chemistry*; D013696:Temperature
D008903:Minerals; D018621:PrPC Proteins; D018620:PrPSc Proteins; D012822:Silicon Dioxide