Rees Helen C, Maddison Ben C, Owen Jonathan P, Whitelam Garry C, Gough Kevin C
Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, UK.
Mol Biotechnol. 2009 Mar;41(3):254-62. doi: 10.1007/s12033-008-9129-5. Epub 2008 Dec 5.
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.
能够沉淀与疾病相关的朊病毒蛋白（PrP(Sc)）的试剂对于开发高灵敏度检测方法以检测生物材料中低水平的这种疾病标志物至关重要。在此，一系列矿物质被证明能沉淀绵羊细胞朊病毒蛋白（PrP(C)）和绵羊瘙痒病PrP(Sc)。二氧化硅沉淀朊病毒蛋白不受PrP(Sc)毒株或宿主物种的影响，该方法可用于沉淀牛海绵状脑病。此方法能可靠地浓缩来自临床感染动物的1.69微克脑蛋白，该脑蛋白稀释于50毫升缓冲液或15毫升血浆中所产生的抗蛋白酶绵羊PrP(Sc)（PrP(res)）。在PrP(res)的免疫学检测中引入二氧化硅沉淀步骤可将检测灵敏度提高1500倍以上。诸如二氧化硅之类的矿物质是容易获得、低成本的试剂，可普遍应用于浓缩与疾病相关的朊病毒蛋白。
