Immunoproteomics and detection of viral diseases

免疫蛋白质组学和病毒性疾病检测

• 批准号: 6792044
• 负责人: MICHAEL D. BOYLE
• 金额: $ 5.83万
• 依托单位: JUNIATA COLLEGE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-15 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6792044
• 关键词: Poxviridae; antiviral antibody; biohazard detection; biotechnology; immunologic assay /test; immunoregulation; interferon gamma; laboratory mouse; mass spectrometry; proteomics; technology /technique development; virus diseases

DESCRIPTION (provided by applicant): Rapid detection of infectious agents has become more relevant with concerns associated with detection of bioterrorist agents. Current technology for detection and quantification of clinically relevant molecules is based on monoclonal and polyclonal antibodies used for immunoassay or detection of nucleic acids using RT-PCR systems. These assays are relatively slow taking at least 3 hours to complete. A novel, rapid, immunoproteomics assay based on high affinity specific antibodies coupled with a mass spectral read out is proposed. In a series of preliminary studies we have demonstrated the ability to use a single antibody-coated antigen capture and transfer reagent (ACTR) to detect model antigens, with good sensitivity, using surface enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF). This immunoproteomic assay can be complete in < 1 hour and follows a sequential process in which antibody is immobilized in step 1, antigen is captured in step 2 and bound antigen is analyzed by mass spectrometry in step 3. Immunoproteomic assays may potentially be faster and as sensitive as the current antibody assays and could provide evidence for bacterial or viral antigens or specific antibodies to these products in samples for an infected animal. This RO3 proposal is designed to optimize this platform technology. In the initial proof-of-concept studies, we will develop and evaluate immunoproteomic assays to measure INF gamma (Th1 cytokine) and specific anti-viral antibodies in the serum of ectromelia infected mice. Ectromelia is a surrogate of variola virus, a CDC Category A bioterrorist agent and thus, the successful immunoproteomic approach may be relevant for detection of bioterrorist agents.

描述（由申请人提供）：传染性病原体的快速检测与生物恐怖主义制剂的检测越来越相关。目前用于检测和定量临床相关分子的技术是基于用于免疫测定的单克隆和多克隆抗体或使用RT-PCR系统检测核酸。这些检测相对较慢，至少需要3小时才能完成。提出了一种新的、快速的、基于高亲和力特异性抗体的免疫蛋白质组学分析方法。在一系列初步研究中，我们已经证明了使用单一抗体包被抗原捕获和转移试剂（ACTR）检测模型抗原的能力，使用表面增强激光解吸电离飞行时间质谱（SELDI-TOF）具有良好的灵敏度。该免疫蛋白质组学分析可在1小时内完成，并遵循以下顺序过程：第1步固定抗体，第2步捕获抗原，第3步通过质谱分析结合抗原。免疫蛋白质组学分析可能会比目前的抗体分析更快、更敏感，并且可以为受感染动物样本中的细菌或病毒抗原或这些产物的特异性抗体提供证据。本RO3提案旨在优化该平台技术。在最初的概念验证研究中，我们将开发和评估免疫蛋白质组学分析，以测量感染鼠血清中的INF γ （Th1细胞因子）和特异性抗病毒抗体。蜱虫是美国疾病控制与预防中心a类生物恐怖制剂天花病毒的替代物，因此，成功的免疫蛋白质组学方法可能与生物恐怖制剂的检测有关。