New method for detection of prion diseases

检测朊病毒病的新方法

• 批准号: 6695039
• 负责人: Andrzej K Drukier
• 金额: $ 10万
• 依托单位: BIOTRACES, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6695039
• 关键词: biomarker; diagnosis design /evaluation; diagnostic tests; differential display technique; electromagnetic radiation; high throughput technology; human tissue; image enhancement; immune tolerance /unresponsiveness; immunologic assay /test; mass spectrometry; microarray technology; neuritic plaques; posttranslational modifications; prions; protein quantitation /detection; protein sequence; proteomics; receptor expression; recombinant proteins; spongiform encephalopathy

DESCRIPTION (provided by applicant): We propose new methods for discovery of TSE biomarkers based on combing the MultiPhoton Detection (MPD) [1,2,3,4] technique with methods of proteomics [5,6,7,8], especially by applying MPD enhanced differential display of proteins (dd-PROT/MPD) and immunoassay (IA/MPD). Subsequently, the methods will be extended to MPD enhanced supersensitive protein chips (P-chips/MPD). These MPD enhanced techniques, developed by the group of Dr. A. K. Drukier, enable more sensitive and less expensive tests for correlating the levels of low abundance protein ("molecular switch") with TSE infections. These differentially displayed proteins will then be validated as TSE biomarkers. The primary objective of Phase I of this proposal is to discover differentially displayed proteins that are down regulated in TSE cases. A similar study was conducted a few years ago by Dr. M. Harrington, on CJD patients. This pioneering study was performed using silver stained 2D gels and revealed those differentially displayed proteins present at relatively high levels (1-10 fmole/ml) in CSF. Only one clearly up-regulated protein and a few possibly down-regulated proteins were discovered. The new tools of proteomics, such as dd-PROT/MPD and high-sensitivity mass spectroscopy, may allow detection and identification at the few attomole/ml level. Our study using proteomics methods sensitive to attomole/ml levels is expected to detect a few tens of differentially expressed proteins. Hopefully, some of them will also be present in blood to allow ante mortem TSE diagnosis. The identification of proteins via genomic information permits the synthesis of recombinant proteins and development of appropriate monoclonal antibodies (mAbs). If the importance of several TSE biomarkers is confirmed, we will develop dedicated supersensitive P-chips (P-chips/MPD) for quantifying up to 64 targeted differentially displayed proteins at 50 fg/ml. Thus, low cost ante mortem TSE detection methods will be developed.

描述（由申请人提供）：我们提出了基于多光子检测（MPD）[1，2，3，4]技术与蛋白质组学方法[5，6，7，8]相结合的发现TSE生物标志物的新方法，特别是通过应用MPD增强的蛋白质差异显示（dd-PROT/MPD）和免疫测定（IA/MPD）。随后，该方法将被扩展到MPD增强的超灵敏蛋白芯片（P-chips/MPD）。这些MPD增强技术是由A. K. Drukier，使更敏感和更便宜的测试相关的低丰度蛋白质（“分子开关”）的水平与TSE感染。然后将这些差异显示的蛋白质验证为TSE生物标志物。 该提案的第一阶段的主要目标是发现在TSE病例中下调的差异显示蛋白。几年前，M.哈灵顿，关于克雅二氏症患者。该开创性研究使用银染色的2D凝胶进行，并揭示了在CSF中以相对高的水平（1-10 fmole/ml）存在的那些差异显示的蛋白质。只有一个明显上调的蛋白质和一些可能下调的蛋白质被发现。蛋白质组学的新工具，如dd-PROT/MPD和高灵敏度质谱，可能允许在几个阿托摩尔/毫升水平上进行检测和鉴定。我们的研究使用蛋白质组学方法敏感的阿托摩尔/毫升水平，预计将检测几十个差异表达的蛋白质。希望它们中的一些也会存在于血液中，以允许生前TSE诊断。 通过基因组信息识别蛋白质可以合成重组蛋白质并开发适当的单克隆抗体（mAb）。如果几种TSE生物标志物的重要性得到证实，我们将开发专用的超灵敏P-芯片（P-芯片/MPD），用于在50 fg/ml下定量多达64种靶向差异显示蛋白。因此，将开发低成本的死前TSE检测方法。