Epitope Discovery via Nanocluster Presentation

通过纳米簇展示发现表位

基本信息

批准号：
7783751
负责人：
DAVID E CLIFFEL
金额：
$ 29.65万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2012-02-28
项目状态：
已结题

项目摘要

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.

该提案的目的是开发一个多功能纳米颗粒平台，用于表位呈现金纳米簇与免疫学分析，用于创建针对免疫的纳米技术系统。纳米颗粒疫苗的未来发展需要蛋白质的产生忠实地再现病原微生物保护性抗原构象的结构。讽刺的是，我们逻辑设计具有保护性的重组或合成蛋白质抗原的能力是相当强的。有限的。由于天然蛋白质上的大多数表位都可能被组装，因此最重要的表位之一表位测定工作面临的挑战是鉴定构象组装的表位。我们假设目标蛋白环结构在金纳米团簇表面的呈现将呈现潜在环构象统计上显着的多样性，为发现致病毒素未知表位的新方法。为了论证这个方案的可行性方法，我们正在创建一个表位标记的纳米簇，模拟芽孢杆菌的保护性抗原（PA）炭疽病。在本提案中，我们的目标是完善和扩展我们的表位概念策略通过以下具体目标介绍纳米团簇： 1.金表面线性表位和构象表位的设计、合成和组装纳米团簇。 2. 确定哪种构象肽环结构忠实地代表了天然存在的肽环结构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.优化多功能纳米粒子在小鼠体内的免疫反应。纳米技术的具体挑战将是在纳米粒子上创建特定的子结构表面，这些特定构象的免疫识别，多功能纳米颗粒的创建，以及肽环结构与纳米颗粒尺寸和曲率函数的相关性。具体变化为生物学和医学包括未映射的线性和构象表位的发现、免疫对功能化纳米粒子的系统响应，筛选抗体库的构象表位，以及最终开发构象表位标记的纳米粒子作为潜在的疫苗。