Structural characterization and proteomic analysis of outer membrane vesicles from Pasteurella multocida strain p52

Abstract

Outer membrane vesicles (OMVs) are bacterial-origin natural nanoparticles potentially emerging as vaccine adjuvants for different Nano-vaccine formulations. They are the spherical derivative product of pathogenic or nonpathogenic bacteria including both gram negative and gram positive. Some archaea and eukaryotes like fungi also generate OMVs which may possess inherent immune-stimulatory properties and can act as delivery candidates for different cargo proteins. In this study, we report the production and characterization of bacterial OMVs collected from vaccine strain P52 of P. multocida and discuss their role as adjuvants in vaccine development. Brain heart infusion broth (BHI) with yeast extract was used to grow Pasteurella multocida. Using ultracentrifugation procedure OMVs were isolated and characterized by different protocols viz Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), Zeta potential, SDS-PAGE, and LC-MS/MS. The collected OMVs with a protein concentration of 8.7 mg/ml were spherical, bilayer with size of 75 to 300 nm in diameter. Proteomic analysis revealed the presence of a group of proteins like the major outer membrane protein and OMP A, which may cause the production of robust immune responses. The findings related to OMV of P. multocida provide foresight in the field of adjuvants and novel vaccine development.