Outer Membrane Vesicles (OMVs) and Proteomics

摘要 外膜囊泡 (OMV) 和蛋白质组学核心 (Core B) 的总体目标是提供 标准化 OMV 制剂的集中平台；高纯度抗原；以及蛋白质组学和生物信息学 分析以加强淋病疫苗研究中心 (GV CRC) 的活动。核心B是关键 GV CRC 的综合组成部分，将为所有四个研究项目和主办方提供服务 响应监控核心（核心 C）。为了成功实现核心 B 的目标，集中化验已 具体分为四个目标： 1）提供OMV：我们自然会提供大量标准化的 详细阐述并提取 OMV，用于所有项目中的统一疫苗接种研究。我们将确保高质量 通过使用优化的生长条件、亚细胞分级、传输来保证所有 OMV 的一致性 电子显微镜和包含 20 种不同蛋白质抗体的免疫印迹 Sikora 实验室开发的标记； 2）纯化抗原：我们将提供纯正完整的抗原，具有天然的 构象保留，使用优质试剂，确保实验一致； 3) 进行蛋白质组学 对于淋病疫苗：我们将进行亚细胞分离、样品制备和处理、高 用于全面抗原挖掘的吞吐量定量蛋白质组学和免疫蛋白质组学研究 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，对所有项目的蛋白质组数据集进行挖掘和广泛分析 生物信息学研究。我们还将准备要提交的数据集并将数据公开存储 可用的数据库。 Core B 对 GV CRC 极其有益，因为它可以实现不同实验室之间的一致研究 并以高效且具有成本效益的方式促进研究进展。

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.