Epitope Discovery via Nanocluster Presentation

通过纳米簇展示发现表位

• 批准号: 7026629
• 负责人: DAVID E CLIFFEL
• 金额: $ 28.24万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7026629
• 关键词: Bacillus anthracis; antibody; antigen antibody reaction; antigen presentation; biomimetics; bioterrorism /chemical warfare; clinical research; conformation; diagnosis design /evaluation; epitope mapping; genetic library; gold; human tissue; immunologic techniques; immunotoxicity; laboratory mouse; microarray technology; nanotechnology; peptide chemical synthesis; surface property; technology /technique development

DESCRIPTION (provided by applicant): The purpose of this proposal is to develop a multifunctional nanoparticle platform for epitope presentation on gold nanoclusters with immunological analysis for the creation of nanotechnologies that target the immune system. The future development of nanoparticle-based vaccines requires the generation of protein structures that faithfully recapitulate the conformation of protective antigens of pathogenic microorganisms. Ironically, our ability to logically design recombinant or synthetic protein antigens that are protective is quite limited. As most epitopes on native proteins are likely to be assembled, one of the most significant challenges facing epitope determination efforts is the identification of conformationally assembled epitopes. We hypothesize that the presentation of target protein loop structures on the surface of gold nanoclusters will present a statistically significant diversity of potential loop conformations to provide the foundation of a new method for discovering unknown epitopes of pathogenic toxins. To demonstrate the feasibility of this approach, we are creating an epitope-labeled nanocluster that mimics the protective antigen (PA) of Bacillus anthracis. In this proposal, our goal is to refine and extend our conceptual strategies for epitope presentation on nanoclusters via the following specific aims: 1. Design, synthesize, and assemble linear and conformational epitopes on the surface of gold nanoclusters. 2. Determine which of the conformational peptide loop constructs faithfully represent the naturally occurring immunogenic structures in native protein. 3. Screen mouse and human antibody libraries, and patient sera using epitopes presented on nanoclusters working towards the development of a diagnostic microarray. 4. Optimize the immunological response of multifunctional nanoparticles in mice. Specific challenges in nanotechnology will be the creation of specific substructures on a nanoparticle surface, immunorecognition of these specific conformations, creation of multifunctional nanoparticles, and correlation of peptide loop structure as a function of nanoparticle size and curvature. Specific changes in biology and medicine include the discovery of unmapped linear and conformational epitopes, the immune system response to functionalized nanoparticles, the screening of antibodies libraries for conformational epitopes, and ultimate development towards conformational epitope-labeled nanoparticles as potential vaccines.

描述（由申请人提供）：该提案的目的是开发一个多功能纳米颗粒平台，用于对金纳米群的表位呈现，并通过免疫学分析来创建针对免疫系统的纳米技术。基于纳米颗粒的疫苗的未来发展需要忠实地概括致病微生物保护性抗原的构象的蛋白质结构。具有讽刺意味的是，我们在逻辑上设计具有保护性的重组或合成蛋白抗原的能力非常有限。由于大多数对天然蛋白质上的表位可能会组装在一起，因此表位确定工作的最重要的挑战之一是识别构型组装的表位。我们假设在金纳米群体表面的靶蛋白环结构的呈现将呈现潜在环构象的统计学意义多样性，以提供一种新方法，以发现发现未知的致病毒素表位。为了证明这种方法的可行性，我们正在创建一个表位标记的纳米簇，该纳米簇模仿炭疽芽孢杆菌的保护性抗原（PA）。在此提案中，我们的目标是通过以下特定目的在纳米群体上提出并扩展我们的表位概念策略：1。设计，合成和组装金纳米群落表面上的线性和构象表位。 2。确定哪种构象肽环构建忠实地代表天然蛋白质中天然存在的免疫原性结构。 3。筛查小鼠和人类抗体库，以及使用纳米簇的表位，用于开发诊断微阵列的表位。 4。优化小鼠多功能纳米颗粒的免疫学反应。纳米技术中的具体挑战将是在纳米颗粒表面上创建特定的子结构，对这些特定构型的免疫认知，创建多功能纳米颗粒以及肽循环结构的相关性作为纳米颗粒大小和曲率的函数。生物学和医学的特定变化包括发现未塑造的线性和构象表位，对功能化纳米颗粒的免疫系统反应，筛选构象表位的抗体文库以及构象表位表位标记的纳米颗粒作为潜在疫苗的构象表位标记的纳米颗粒的最终发展。