Cell Mimic Microarrays for the Multivalent Pathogen Profiling & Characterization

用于多价病原体分析的细胞模拟微阵列

基本信息

批准号：
8310935
负责人：
Athena Guo
金额：
$ 36.45万
依托单位：
MICROSURFACES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-03 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8310935
关键词：
Address Adhesions Affinity Air Antigens Area Avian Influenza Avidity Binding Biological Models CD209 gene Cell membrane Cell surface Cells Chemicals Complex Dendritic Cells Development Disease Outbreaks Ensure Environment Escherichia coli Glycolipids Glycoproteins Goals HIV-1 Hemagglutinin Lead Ligands Link Lipid Bilayers Mannose Methodology Microarray Analysis Mono-S Mutation Nature Pattern Phase Polysaccharides Probability Procedures Rehydrations Research Research Project Grants Role Screening procedure Specificity Spottings Staging Surface Techniques Technology Vaccines Variant Vertebral column Viral Vaccines Virus combat density effective therapy high throughput screening influenzavirus inhibitor/antagonist mannose receptor pathogen receptor receptor binding research and development research study sugar swine flu tool vaccine development vaccine evaluation

项目摘要

DESCRIPTION (provided by applicant): This research project aims to develop fluidic glycan microarrays for the quantitative profiling and characterization of pathogens, and for the screening of pathogen inhibitors and the development of vaccines. The approach targets a common mechanism at the initial stage of pathogen attack: the recognition of and attachment onto host cells via multivalent interaction between receptor proteins and glycan molecules. The tremendous variation in glycans and the complexity in multivalent interaction have necessitated the use of large-scale profiling and analysis techniques, particularly glycan microarrays. The proposed fluidic approach overcomes two major limitations of current glycan microarray technology: the lack of mobility and the difficulty in quantitatively controlling glycan density. Multivalent cell surface interactions often require mobility of the fluidic cell membrane environment and are strong functions of surface glycan density. In order to quantitatively apply the glycan microarray in profiling and characterization, one must ensure mobility and control of glycan density over a broad range. The specific aims during phase-II are: Aim 1: using haemagglutinin, a predominant antigen on influenza viruses, and the dendritic cell receptor DC-SIGN, a binding receptor for mannose moieties on HIV-1 virus, as model systems and establish the roles of secondary interactions in binding affinity, avidity, and specificity. These experiments will establish the general applicability of the fluidic microarrays in profiling and characterizing complex pathogen-cell surface interactions; Aim 2: using several strains of E. coli with varying affinity and selectivity towards mannose as model systems and establishing that the fluidic and density gradient glycan microarray can be used to quantitatively profile the variability in binding affinity and multivalency among strains of the same species. Quantifying such variability is essential to the understanding and surveillance of how random mutations can lead to new pathogen threats, as exemplified by the recent outbreaks of avian flu and swine flu; Aim 3: To establish chemical procedures for the optimization of the fluidic glycan microarray, including spatial confinement of the supported lipid bilayer spots, efficient blocking of surfaces outside the spotted areas, recoverability in drying and rehydration of the microarrays, and long term stability of content glycan microarrays. These practical issues must be addressed in developing the fluidic glycan microarray as a viable product. The long-term goal of this R&D plan is to develop an effective high-throughput tool in the combat against pathogen threats.

描述（由申请人提供）：该研究项目旨在开发流体聚糖微阵列，以进行病原体的定量分析和表征，以及筛查病原体抑制剂和疫苗的发展。该方法在病原体攻击的初始阶段靶向一种常见的机制：通过受体蛋白与聚糖分子之间的多价相互作用的识别和附着在宿主细胞上。聚糖的巨大变化以及多价相互作用的复杂性需要使用大规模的分析和分析技术，尤其是聚糖微阵列。提出的流体方法克服了当前聚糖微阵列技术的两个主要局限性：缺乏迁移率和定量控制聚糖密度的困难。多价细胞表面相互作用通常需要流体细胞膜环境的迁移率，并且是表面聚糖密度的强大功能。为了定量地将聚糖微阵列应用于分析和表征，必须确保在广泛范围内的迁移率和对聚糖密度的控制。第三阶段期间的具体目的是：目标1：使用血凝素（一种主要的流感病毒抗原）和树突状细胞受体DC-SIGN，一种在HIV-1病毒上的甘露糖基部分的结合受体作为模型系统，并确定二级相互作用在结合亲属亲属和特定性的二级相互作用中的作用。这些实验将在分析和表征复杂的病原体表面相互作用方面建立流体微阵列的一般适用性； AIM 2：使用具有不同亲和力和对甘露糖的选择性的几种大肠杆菌作为模型系统的选择性，并确定可以使用液体和密度梯度梯度聚糖微阵列来定量介绍同一物种菌株之间结合亲和力和多价性的可变性。量化这种变异性对于理解和监视随机突变如何导致新病原体威胁至关重要，这是近期禽流感和猪流感爆发的例证。目的3：建立化学程序来优化流体聚糖微阵列，包括对受支持的脂质双层斑点的空间限制，有效地阻断斑点区域外的表面，在斑点区域内的可恢复性，在微阵列的干燥和再渗透中的可恢复性以及内容的长期稳定性的是Glycan Microarays的稳定性。在开发流体聚糖微阵列作为可行的产品时，必须解决这些实际问题。该研发计划的长期目标是在针对病原体威胁的战斗中开发有效的高通量工具。