Immunoproteomics and detection of viral diseases

描述（由申请人提供）：传染性病原体的快速检测与生物恐怖分子病原体检测相关的问题更加相关。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或使用RT-PCR系统检测核酸的单克隆和多克隆抗体。这些测定相对较慢，至少需要3小时才能完成。提出了一种新的，快速的，免疫蛋白质组学分析的基础上，高亲和力的特异性抗体与质谱读出耦合。在一系列的初步研究中，我们已经证明了使用单一抗体包被的抗原捕获和转移试剂（ACTR）检测模型抗原的能力，具有良好的灵敏度，使用表面增强激光解吸电离飞行时间质谱（SELDI-TOF）。该免疫蛋白质组学测定可以在< 1小时内完成，并且遵循顺序过程，其中在步骤1中固定抗体，在步骤2中捕获抗原，并且在步骤3中通过质谱分析结合的抗原。免疫蛋白质组学检测可能比目前的抗体检测更快、更灵敏，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品的特异性抗体提供证据。该RO 3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测定法，以测量感染小鼠血清中的INF γ（Th 1细胞因子）和特异性抗病毒抗体。鼠痘是天花病毒的替代物，天花病毒是CDC A类生物恐怖分子制剂，因此，成功的免疫蛋白质组学方法可能与生物恐怖分子制剂的检测相关。

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.