Betz, ElizabethPlavchak, ChristineHansen, RileyWilliams, Kim R.2023-11-212023-11-21http://hdl.handle.net/11124/178558https://doi.org/10.25676/11124/178558Extracellular vesicles (EVs) are nano-vesicles (50-500 nm) that are released by cells and play an important role in cell-to-cell communication. EVs have applications ranging from disease treatment and detection to developing more efficient biofuels. Outer membrane vesicles (OMVs) are a specific type of EV released by gram-negative bacteria. Specifically, OMVs from the soil bacteria Pseudomonas putida, contain enzymes with the ability to break down lignin, a complex plant polymer that is a plentiful resource for biofuels. Studying the relationship of size and content of OMVs within a cultivation of vesicles aids in the understanding of how these OMVs efficiently break down lignin. To investigate P. putida OMVs, we hypothesized that asymmetrical flow field-flow fractionation (AF4), dynamic light scattering (DLS), and multi-angle light scattering (MALS) could be used to efficiently separate and determine the size distributions of OMVs. Liposome standards were utilized in initial experiments to analyze retention times according to size due to their similar structure to OMVs. AF4-DLS-MALS fractionations of P. putida OMVs showed two different sized populations when grown on lignin. Separating these OMV populations will help correlate differences in protein and lipid content between small (50-150 nm) and large (200-500 nm) vesicles.articlesengCreative Commons CC-BY License or the Creative Commons CC-BY-NC License.outer membrane vesiclesasymmetrical flow field-flow fractionationsize distributionsText