Levels. Summary/Conclusion: CH promotes EV release from HepG2 cells. EV from hypoxic FFA-treated HepG2 cells evoke pro-fibrotic responses in LX-2 cells. Further genomic and proteomic characterization of EV released by steatotic cells below hypoxia are required to further delineate their function inside the crosstalk in between hepatocytes and stellate cells within the setting of NAFLD and OSAS. Funding: FONDECYT 1150327150311.Helmholtz-Institute for Pharmaceutical Study Saarland, Biogenic Nanotherapeutics, Saarbruecken, Germany; bHelmholtz-Institute for Pharmaceutical Analysis Saarland, Drug Design and Optimization, Saarbruecken, Germany; 3Helmholtz-Institute for Pharmaceutical Investigation Saarland, BION, Saarbruecken, GermanyIntroduction: Introducing bacteria-binding modest molecules towards the surface of outer membrane vesicles (OMVs) could greatly enhance their prospective for antimicrobial drug delivery also hard to treat bacteria. Among the small number of studies on surface modification of OMVs, extremely handful of deal with little molecules. The aim on the present study is to evaluate different techniques of introducing bacteria specific targeting moieties to OMVs. We assessed the modification of surface proteins utilizing Nhydroxysuccinimide (NHS) esters, well established for mammalian extracellular vesicles (EVs), cholesterol insertion, primarily applied for liposomes, and the novel application of diazo-transfer followed by click-chemistry. Approaches: OMVs had been obtained from model myxobacteria by differential ultracentrifugation (UC) followed by size-exclusion chromatography (SEC). For cholesterol insertion and NHS ester-modification, purified OMVs were incubated with either cholesteryl PEG 2,000 FITC or sulfo cyanine7 NHS ester. For diazo transfer the pellet right after UC was incubated with a diazo transfer agent and the OMVs subsequently conjugated with DBCO-AF594. Unincorporated dye was removed by SEC. Liposomes had been composed of DMPC and DPPC in two:3 molar ratio. Results represent correlated fluorescence intensity and particle quantity. Benefits: Therapy with sulfo cyanine7 NHS ester led towards the modification with 547 163 molecules per OMVs, when CD233 Proteins Accession compared with 18 1 for the manage working with sulfo cyanine7 acid. Cholesterol insertion introduced 4 1 molecules per OMV, when compared with 101 23 for liposomes. Initial outcomes for the diazo-transfer showed 71 dye-molecules per OMV, with 32 for the control. Summary/Conclusion: From the three strategies, NHS ester-modification displayed the highest efficiency, comparable to published benefits for mammalian EVs. In comparison, diazo transfer only yielded 13 in the dye-molecules per particle. On the other hand, you will discover nevertheless lots of parameters to be optimized for this technique, which includes OMV concentration and incubation period. Cholesterol insertion was unsuccessful for OMVs,ISEV2019 ABSTRACT BOOKprobably owing to their membrane structure. Within this study, we aim to have crucial insights into the modification of OMVs for bacterial targeting and EV-surface engineering generally. Funding: This project was funded by Studienstiftung des Deutschen Volkes and Bundesministerium fuer Bildung und Forschung.OWP1.09=LBT01.CEACAM1 Proteins Biological Activity Coagulation influences properties of extracellular vesicles isolated from autologous blood derived solutions Andrea De Lunaa, Alexander Otahala, Olga Kutenb, Zsombor Laczac and Stefan NehreraaDanube University Krems, Krems, Austria; bOrthoSera GmbH, Krems, Austria; cOrthosera GmbH, Krems, AustriaOWP1.08=LBT02.Isolation of neuron-specific extracellular vesicles Dmitr.