Outer Membrane Vesicles (OMVs) and Proteomics

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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.