D resulting in a loss ofISEV2019 ABSTRACT BOOKbead fluorescence that may be measured making use of highthroughput flow cytometry. These biosensors were assayed applying either recombinant proteinases or isolated EVs from in vitro cancer models. Results: Human metalloproteinase recognition motifs had been identified MSR1/CD204 Proteins Recombinant Proteins inside the literature along with a total of 70 various metalloproteinase biosensors have been created. A handle biosensor (PhaC-112L-T-G) detected 0.five U of tobacco etch virus protease (AcTEV) activity along with the PhaC-112L-P14-G biosensor, in spite of some background off-target activity, was in a position to detect 0.033 mU of recombinant MMP14 activity. Membrane-bound metalloproteinases MMP14 and ADAM10 had been also detected in EVs isolated (ultracentrifugation) from in vitro cancer models. Summary/Conclusion: Our biosensors detected EVassociated metalloproteinases and could serve as beneficial research tools for EV-biomarker discovery. Funding: Dr Richard Kelwick is funded by a Royal Society of Edinburgh Enterprise Fellowship and an Imperial Self-confidence in Idea 2018 grant. We also acknowledge the help of Engineering and Physical Science Investigation Council (EPSRC) grants [EP/ L011573/1; EP/P028519/1] plus the Biotechnology and Biological Sciences Analysis Council (BBSRC) Foundry grant [BB/L027852/1].resolution imaging on the identical device. Specifically, the surface of your imaging chamber is passivated with anti-CD 63 to capture the DiD stained vesicles. The acquisition on the raw image series was accomplished using total internal reflection fluorescence PSGL-1/CD162 Proteins Recombinant Proteins microscopy (TIRF) having a 642-nm diode laser for excitation. Two forms of super-resolution procedures were tested which includes super resolution radial fluctuations (SRRF) and stochastic optical reconstruction microscopy (STORM). Benefits: The size of single exosomes within the final pictures have been estimated by the full-width at half-maximum (FWHM) of Gaussian fitted to the distribution of single molecules. We have located that the resolution limit in the single particle is lowered to 70 nm. The preliminary information from SRRF and STORM showed the particle size and size distribution had been compared to nanoparticle tracking evaluation (NTA) benefits. Summary/Conclusion: This system provides in-depth size evaluation of single exosomes beneath the diffraction limit. On top of that, capturing exosomes from coarsely isolated samples by way of specific antibodies would reduce the time needed for sequential ultracentrifugation, the present typical method for exosome isolation. Finally, this imaging chamber presents a versatile platform for protein profiling as the captured exosomes is often labelled with distinct antibody-dye conjugates to reveal the surface proteins contents.PT09.Single exosome size analysis utilizing super resolution microscopy Xia Lia, Mina Hoorfarb and Isaac Liaa University of British Columbia Okanagan, Kelowna, Canada bDepartment of Chemistry, University of British Columbia Okanagan, Kelowna, CanadaPT09.12=OWP3.Identification of single tumour-derived extracellular vesicles by suggests of optical tweezers and Raman spectroscopy Agustin Enciso-Martineza, Edwin van der Polb, Aufried Lenferinkc, Leon Terstappena and Cees Ottoa Medical Cell Biophysics, University of Twente, Enschede, Netherlands; Amsterdam UMC, University of Amsterdam, Department of Biomedical Engineering and Physics, Amsterdam, Amsterdam, Netherlands; cUniversity of Twente, Enschede, Netherlandsb aIntroduction: Exosomes are a sort of extracellular vesicle (EV) with diameters of 3050 nm and are s.