Outer Membrane Vesicles (OMVs) and Proteomics

外膜囊泡 (OMV) 和蛋白质组学

• 批准号: 9900721
• 负责人: Aleksandra Elzbieta Sikora
• 金额: $ 25.98万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9900721
• 关键词: Antibodies; Antigens; Area; Bioinformatics; Biological Assay; Cell Fractionation; Data; Data Analyses; Data Set; Databases; Deposition; Ensure; Epitopes; Goals; Growth; Immune response; Immunization; Immunoblotting; 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; Work; cell envelope; cost effective; experimental study; gonorrhea vaccine; preservation; product development; protein biomarkers; 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.

摘要 外膜囊泡（OMV）和蛋白质组学核心（核心B）的总体目标是提供一个 标准化OMV制剂的集中平台;高纯度抗原;蛋白质组学和生物信息学 加强淋病疫苗研究中心（GV CRC）的活动。核心B是一个枢纽 GV CRC的综合组成部分，并将为所有四个研究项目和东道主提供服务 响应监测核心（核心C）。为了成功实现核心B的目标，集中式分析 我们分为四个具体目标：1）提供OMV：我们将提供大量的标准化，自然 为所有项目的统一疫苗接种研究制定和提取OMV。我们将确保高品质 通过使用优化的生长条件、亚细胞分级分离、传输 电子显微镜和免疫印迹，包括20种不同的抗蛋白质的抗体阵列 Sikora实验室开发的标记物; 2）纯化抗原：我们将提供纯的和完整的抗原， 构象保留，使用高质量的试剂，以确保一致的实验; 3）进行蛋白质组学 淋病疫苗：我们将进行亚细胞分离，样品制备和处理，高- 通量定量蛋白质组学和免疫蛋白质组学研究，用于全面的抗原挖掘， 定义OMV内容; 4）采用综合生物信息学平台。我们将提供抗原表位 通过广泛的下游应用，为所有项目挖掘和广泛分析蛋白质组学数据集 生物信息学研究我们还将准备提交的数据集，并将数据存款公开 可用的数据库。 核心B对GV CRC非常有利，因为它可以在不同实验室之间进行一致的研究 并以有效和具有成本效益的方式促进研究进展。