Outer Membrane Vesicles (OMVs) and Proteomics

外膜囊泡 (OMV) 和蛋白质组学

• 批准号: 10362590
• 负责人: Aleksandra Elzbieta Sikora
• 金额: $ 21.75万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-26 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10362590
• 关键词: Antibodies; Antigens; Area; Bioinformatics; Biological Assay; Cell Fractionation; Data; Data Analyses; Data Set; Deposition; Ensure; Epitopes; Goals; Growth; Immune response; Immunization; Investigation; Laboratories; Membrane; Methodology; Mining; Molecular Conformation; Monitor; Neisseria gonorrhoeae; Neisseria meningitidis; Peptides; Preparation; Process; Proteomics; Reagent; Recombinants; Research; Research Personnel; Research Project Grants; Sampling; Services; Standardization; Transmission Electron Microscopy; Vaccine Research; Vesicle; Western Blotting; Work; cell envelope; cost effective; experimental study; gonorrhea vaccine; preservation; product development; protein biomarkers; public database; vaccine trial

ABSTRACT The overall objective of the Outer Membrane Vesicles (OMVs) and Proteomics Core (Core B) is to provide a centralized platform for standardized OMV preparations; highly pure antigens; and proteomics and bioinformatics analyses to enhance the activities of the Gonorrhea Vaccine Research Center (GV CRC). Core B is a pivotal integrative component of the GV CRC and will provide services for all four research projects and the Host Response Monitoring Core (Core C). To successfully accomplish the goals of Core B, centralized assays have been divided into four specific aims: 1) Provide OMVs: We will provide large quantities of standardized, naturally elaborated and extracted OMVs for uniform vaccination studies in all projects. We will ensure the high quality and consistency of all OMVs by using optimized growth conditions, subcellular fractionation, transmission electron microscopy, and immunoblotting with an inclusive array of 20 different antibodies against protein markers developed by the Sikora Lab; 2) Purify Antigens: We will provide pure and intact antigens, with native conformation preserved, using high quality reagents to ensure consistent experiments; 3) Conduct Proteomics for Gonorrhea Vaccines: We will conduct subcellular fractionation, sample preparation and processing, high- throughput quantitative proteomics, and immunoproteomics studies for comprehensive antigen mining and to define OMV contents; 4) Employ a Comprehensive Bioinformatics Platform. We will provide antigen epitope mining and extensive analyses of proteomics datasets for all projects by applying wide-ranging downstream bioinformatics investigations. We will also prepare the datasets for submission and deposit the data to publicly available databases. Core B is exceedingly beneficial to the GV CRC as it enables consistent studies between different laboratories and enhances research progress in an efficient and cost-effective manner.

抽象的 外膜囊泡（OMV）和蛋白质组学核心（核心B）的总体目标是提供 标准化OMV准备工作的集中式平台；高度纯抗原；以及蛋白质组学和生物信息学 分析以增强淋病疫苗研究中心（GV CRC）的活性。核心B是关键 GV CRC的集成部分，并将为所有四个研究项目和主持人提供服务 响应监视核心（核心C）。为了成功实现核心B的目标，集中测定 被分为四个具体目标：1）提供OMV：我们将提供大量标准化，自然 在所有项目中详细阐述和提取的OMV进行均匀的疫苗接种研究。我们将确保高质量 通过使用优化的生长条件，亚细胞分级，传播，所有OMV的一致性 电子显微镜和具有20种不同抗体的包含阵列的免疫印迹 Sikora Lab开发的标记； 2）纯化抗原：我们将提供纯净和完整的抗原 使用高质量试剂保留构象，以确保一致的实验； 3）进行蛋白质组学 对于淋病疫苗：我们将进行亚细胞分馏，样品制备和加工，高 吞吐量定量蛋白质组学和用于综合抗原开采的免疫蛋白质组学研究 定义OMV内容； 4）采用全面的生物信息学平台。我们将提供抗原表位 通过在下游应用广泛的范围，对所有项目的蛋白质组学数据集进行了挖掘和广泛的分析 生物信息学调查。我们还将准备提交数据集并将数据存放到公开 可用数据库。 核心B对GV CRC非常有益，因为它可以在不同实验室之间进行一致的研究 并以有效且具有成本效益的方式增强了研究进展。