Outer Membrane Vesicles (OMVs) and Proteomics

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 标准化OMV制备的集中平台；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)活动的分析。核心B是一个关键 GV CRC的综合组成部分，将为所有四个研究项目和东道国提供服务 响应监控核心(核心C)。为了成功实现核心B的目标，集中式检测具有 具体目标分为四个：1)提供OMV：我们将提供大量标准化、自然而然的 制定和提取OMV，用于所有项目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、亚细胞分离、传递 电子显微镜和免疫印迹，包括20种不同的蛋白质抗体 由西科拉实验室开发的标记物；2)纯化抗原：我们将提供纯净和完整的抗原，具有天然的 保持构象，使用高质量的试剂确保实验的一致性；3)进行蛋白质组学 对于淋病疫苗：我们将进行亚细胞分离、样品制备和加工，高效 吞吐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和 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学数据集进行挖掘和广泛分析 生物信息学调查。我们还将准备数据集以供提交，并将数据存放到公众 可用的数据库。 核心B对GV CRC非常有利，因为它能够在不同实验室之间进行一致的研究 并以高效和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.