An aggregation-specific enzyme-linked immunosorbent assay:: Detection of conformational differences between recombinant PrP protein dimers and PrPSc aggregates
An aggregation-specific enzyme-linked immunosorbent assay:: Detection of conformational differences between recombinant PrP protein dimers and PrPSc aggregates
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DOI：
10.1128/jvi.79.19.12355-12364.2005
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发表时间：
2005-10-01
影响因子：
5.4
通讯作者：
Sy, MS
中科院分区：
文献类型：
--
作者：
Pan, T;Chang, BG;Sy, MS
正常细胞朊病毒蛋白 PrPC 转化为抗蛋白酶的瘙痒病 PrPSc 聚集体是朊病毒疾病的原因。我们通过筛选 30 种抗 PrP 单克隆抗体 (MAb) 与重组小鼠、绵羊、牛或人 PrP 二聚体反应的能力，开发了一种新型酶联免疫吸附测定 (ELISA)，该测定对 PrP 聚集体具有特异性。一种与所有四种重组 PrP 二聚体反应的 MAb 也与 ME7、139A 或 22L 感染的小鼠大脑中的 PrPSc 聚集体反应。 PrPSc 聚集体具有蛋白酶 K 抗性，质量为 2,000 kDa 或更大，并且在检测不到蛋白酶抗性 PrP 时出现。这种简单而灵敏的检测方法为开发其他物种朊病毒疾病的诊断测试奠定了基础。最后，我们开发的聚集体特异性ELISA的原理可能适用于由异常蛋白质聚集引起的其他疾病，例如阿尔茨海默病或帕金森病。
The conversion of the normal cellular prion protein, PrPC, into the protease-resistant, scrapie PrPSc aggregate is the cause of prion diseases. We developed a novel enzyme-linked immunosorbent assay (ELISA) that is specific for PrP aggregate by screening 30 anti-PrP monoclonal antibodies (MAbs) for their ability to react with recombinant mouse, ovine, bovine, or human PrP dimers. One MAb that reacts with all four recombinant PrP dimers also reacts with PrPSc aggregates in ME7-, 139A-, or 22L-infected mouse brains. The PrPSc aggregate is proteinase K resistant, has a mass of 2,000 kDa or more, and is present at a time when no protease-resistant PrP is detectable. This simple and sensitive assay provides the basis for the development of a diagnostic test for prion diseases in other species. Finally, the principle of the aggregate-specific ELISA we have developed may be applicable to other diseases caused by abnormal protein aggregation, such as Alzheimer's disease or Parkinson's disease.