The production of drug-loaded EVs and to discover possible application for in situ drug delivery technique. Funding: This investigation is funded by Focused Ultrasound Foundation.OS23.Extracellular Vesicles for new Molecular Insight to Biomolecular Interactions Tamas Beke-Somfaia, Priyanka Singhv, Imola Szigyarto and Zoltan VargacaPI, Budapest, Hungary; bMs, Budapest, Hungary; cResearch Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, HungaryIntroduction: The potential of extracellular vesicles (EVs) to revolutionize the diagnosis and therapy of various ailments has been realized and therefore it’s an extensively studied path. However, EVs are also within the size variety appropriate for membrane biophysics, when they preserve the complicated composition of a biological bilayer. Consequently, they are optimal for monitoring the structure, orientation and function of biomolecules connected to EVs.Approaches: The investigated red blood cell-derived vesicles (REVs) have been isolated from blood applying a typical protocol and 4-1BB Inhibitor Storage & Stability purified making use of size-exclusion chromatography. REVs have been subjected to IR, CD and flow-Linear Dichroism spectroscopy, freeze-fracture Transmission Electron Microscopy as well as Dynamic Light Scattering. Outcomes: Right here we demonstrate that polarized light spectroscopy approaches can give important details on REVs and molecules inserting into their bilayer. Flowlinear dichroism (flow-LD) measurements show that EVs could be oriented by shear force, insight into properties of oriented macromolecules in the vesicles. The Soret-band in the LD spectra demonstrates that hemoglobin molecules are oriented and associated for the lipid bilayer in freshly released REVs [1]. Further on, we chosen 3 distinctive antimicrobial peptides (AMPs), CM15, melittin and gramicidin and investigated their interactions with REVs utilizing a diverse set of methods. The peptide-membrane interactions reveal numerous novel function of AMPs, including their capability to take away linked proteins in the surface of REVs (Figure 1). [1] I. Cs. Szigy t R. De , J. Mih y, S. Rocha, F. Zsila, Z. Varga, T. Beke-Somfai. Flow-alignment of extracellular vesicles: structure and orientation of membrane associated biomacromolecules studied with polarized light. ChemBioChem. 2018;19:54551 Summary/Conclusion: In conclusion, EVs offer exceptional opportunities to much better recognize the function and mechanism of organic membrane active biomolecues. Funding: This work was funded by the Momentum programme (LP2016-2), by the National Competitiveness and Excellence Plan (NVKP_16-1-20160007) and BIONANO_GINOP-2.3.2-15-2016-00017. The J os Bolyai Investigation Scholarship (Z.V.) is tremendously acknowledged.JOURNAL OF EXTRACELLULAR VESICLESSymposium Session 24: Mechanisms of EV Delivery Chairs: Pieter Vader; Hang Hubert Yin Place: Level B1, Hall B 13:004:OS24.State with the art microscopy for live cell study of the extracellular vesicle-mediated drug delivery Ekaterina Lisitsynaa, Kaisa Rautaniemia, Heikki Saarib, Timo Laaksonena, Marjo Yliperttulab and Elina Vuorimaa-Laukkanena Laboratory of Chemistry and Bioengineering, Tampere University of Technology, Tampere, Finland; bDivision of Pharmaceutical BioTraditional Cytotoxic Agents Storage & Stability Sciences and Drug Research Program, Faculty of Pharmacy, University of Helsinki, Helsinki, FinlandaSummary/Conclusion: This analysis offers new realtime strategies to investigate EV kinetics with living cells and complements the existing tactics. The findings on the study strengthen the.