Immuno-cryo-EM, gold particles were conjugated with anti-EpCAM or anti-GLPC-1 Abs or Anx52. Benefits: By FC, we discovered that massive amounts of EpCAM+ EVs had been released by CaPan2 and BXPC3 cells, and substantially significantly less (100 by PanC1 and Alpha-1 Antitrypsin 1-4 Proteins Formulation MiaPaCa-2 cells. Additionally, larger amounts ( 4 of PS+ EVs were released by PanC1 and MiaPaCa-2, as compared to CaPan2 and BXPC3 cells. No GPC-1+ EVs had been detected with all the two Abs applied right here inside the four cell lines. By immuno-cryo-EM, we identified EpCAM+ EVs in CaPan2 and BXPC3 supernatants. These EVs ranged in size from 100 nm to 1 . Most EpCAM+ EVs of compact size (100 nm), the so-called exosomes, had been also labelled by Anx5. No labelling was observed together with the antiGPC-1 Abs employed. Western-blotting experiments revealed the presence of GPC-1 in cell extracts for the two Abs. This suggests that GPC-1 proteins are present within the cell cytoplasm, but not in the EV surface. Conclusion: This study supplies a semi-quantitative analysis of EVs secreted by PaCa cell lines, and is at present complemented by a study of EVs present in plasma of PaCa patients. References 1. Melo et al., Nature 2015; 523: 17782. 2. Arraud et al., J. Thromb. Haemost. 2014; 12: 61427. three. Arraud et al., Cytom. A 2016 ; 89: 18495.Introduction: The presence of nucleic acid inside the extracellular vesicles (EVs) has been known for its function in the intercellular communication. The EVs are exfoliated from cells and may be detected in diverse sources such as tissues, blood, urine and stools. Here, we examined the existence of compact RNAs such as miRNAs in EVs with the washed stool from colorectal cancer individuals and analysed them as prospective biomarkers. Approaches: The EV was isolated from washed stool of colorectal cancer patients by utilizing the aqueous two-phase technique (ATPS) and its RNA was purified by treating with TRIzol. The total small RNAs like miRNAs were purified and analysed by modest RNA-seq working with nextgeneration sequencing (NGS) technologies.PF01.Surface glycosylation profiling of evs applying lectin-nanoparticles Parvez Syed1, Laura Lehtinen2, Kamlesh Gidwani1, Khirul Islam3, Janne Leivo4, Kim Pettersson3 and Urpo Lamminm i1Friday, May 19,Division of Biochemistry/Toll-like Receptor 8 Proteins Recombinant Proteins Biotechnology, University of Turku, Finland; Department of Pathology, University of Turku and Turku University Hospital, Turku, Finland; 3Department of Biochemistry, Division of Biotechnology, University of Turku, Finland; 4Department of Urology, Erasmus Medical Centre, Rotterdam, The NetherlandsIntroduction: Extracellular vesicles (EVs) are secreted by almost all cells and present range of proteins, lipids and glycans on their surface. The majority of your surface tumour markers reported to date are either glycoproteins or glycolipids. Traditionally, the EV-surface glycosylation profiling is either carried out making use of mass spectrometry or lectin microarrays. Nevertheless, each these procedures demand isolation of EVs. Within this study, we use lectins, which bind for the glycan part of the glycoproteins, conjugated with Eu3+-doped nanoparticles (NP) to determine the glycans presented around the surface of the EVs. Approaches: The EVs from cell-free cell culture supernatants of HEK293 and ovarian cancer (OvCa) cell lines (SKOV3, M022 and M019i) had been captured applying biotinylated anti-CD63 antibody immobilised onto streptavidin coated 96-well plate. The captured EVs have been probed with lectins conjugated with one hundred nm-sized polystyrene NPs containing 30,000 Eu3+ ions. The lectins utilised in this study had been, galactose binding.