Epitope Discovery via Nanocluster Presentation

这项建议的目的是开发一个多功能的纳米颗粒平台，用于展示表位 具有免疫学分析的金纳米簇，用于创建针对免疫的纳米技术 系统。基于纳米颗粒的疫苗的未来发展需要产生蛋白质 如实概括病原微生物保护性抗原构象的结构。 具有讽刺意味的是，我们逻辑地设计具有保护性的重组或合成蛋白抗原的能力相当强 有限的。由于天然蛋白质上的大多数表位可能是组装的，最重要的是 表位确定工作面临的挑战是识别构象组装的表位。 我们假设靶蛋白质环结构在金纳米团簇表面的呈现 将呈现统计上显著的潜在环状构象的多样性，从而为 发现致病毒素未知表位的新方法。为了证明这一点的可行性 方法，我们正在创建一个模拟芽孢杆菌保护性抗原(PA)的表位标记的纳米簇 炭疽病。在这个提案中，我们的目标是提炼和扩展我们的表位概念策略 通过以下具体目标介绍纳米集群： 1.设计、合成和组装金表面的线性和构象表位 纳米团簇。 2.确定哪一种构象肽环结构忠实地代表自然产生的 天然蛋白质中的免疫原性结构。 3.使用纳米簇上呈现的表位筛选小鼠和人类抗体库，以及患者血清 致力于开发诊断微阵列。 4.优化多功能纳米粒对小鼠的免疫应答。 纳米技术中的具体挑战将是在纳米颗粒上创建特定的亚结构 表面，这些特定构象的免疫识别，多功能纳米颗粒的创建，以及 多肽环结构与纳米颗粒大小和曲率的关系。中的具体变化 生物学和医学包括发现未映射的线性和构象表位，免疫表位 系统对功能化纳米颗粒的反应，构象抗体库的筛选 表位，以及最终向构象表位标记纳米粒子的潜在发展 疫苗。

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 towardsthe 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.