Mon. May 20th, 2024

Ilvia Picciolini1, Alice Gualerzi2, Carlo Contactin-2 Proteins Storage & Stability Morasso2, Renzo Vanna2, Marzia Bedoni3, Massimo Masserini4 and Furio GramaticaLaboratory of Nanomedicine and Clinical Biophotonics CLEC-2 Proteins Formulation LABION, Fondazione Don Gnocchi University of Milano-Biocca, Milano, Italy;Thursday May 18,two Laboratory of Nanomedicine and Clinical Biophotonics LABION, Fondazione Don Gnocchi; 3Laboratory of Nanomedicine and Clinical Biophotonics LABION, Fondazione Don Carlo Gnocchi ONLUS; four University of Milano-Biocca, Milano, ItalyFunding: This function is supported by NWO through a Vidi grant and by STW by way of the Perspectief grant Cancer-ID. (Each to Wouter H. Roos).Introduction: Exosomes have emerged as a brand new class of biomarkers of neurological issues displaying an involvement in neurodegenerative processes. The major interest within this field is supported by the fact that exosomes are in a position to cross the blood brain barrier and can therefore supply the exceptional possibility to study the biochemical processes inside the central nervous program from a biofluid uncomplicated to access as human blood. Inspired by current progresses in plasmonic biosensors that demonstrated their ability to detect exosomes from biological samples, we’ve developed a biosensor primarily based on surface plasmon resonance imaging (SPRi) for the isolation of exosomes of neuronal origin and to study their membrane surface and interactions with distinct biomolecules. Approaches: The SPRi microarray was optimised for the detection of different subpopulations of exosomes extracted by size-exclusion chromatography from plasma of healthful volunteers. Bare gold SPRi chips have been coated using a self assembled monolayer and further activated by EDC/NHS chemistry, in an effort to be functionalised with unique antibodies, deposited by automated microspotting. Soon after exosomes injection on the SPRi chip, we evaluated the interaction involving their membrane molecules and particular antibodies. Results: The surface chemistry was optimised for the immobilisation of antibodies and we tested simultaneously unique antibodies including CD9 and CD63 which might be generic exosomes markers, and CD171/L1 as neuronal marker. As soon as the exosomes had been adsorbed on the chip, the injection of other antibodies was followed by a signal in correspondence of certain exosomes subpopulations, demonstrating the possibility to characterise exosome membranes using a sandwich method. Conclusion: These results recommend that the use of SPRi can assist to simultaneously discriminate and immobilise various exosomes subpopulations and to evaluate the interaction with biomolecules, with a perspective of investigating biological role of those biomarkers.PT05.Detection and characterisation of exosomes in TEM photos utilizing ExosomeAnalyzer: a novel software tool Anna Kotrbov, Karel Stpka2, Martin Maska2, Jakub Jozef P enik2, Ladislav Ilkovics3, Dobromila Klemov, Ales Hampl3, V zslav Bryja1, Vendula Posp halov and Pavel Matula2 Department of Experimental Biology, Faculty of Science, Masaryk University, Czech Republic; 2Centre for Biomedical Image Evaluation, Faculty of Informatics, Masaryk University, Czech Republic; 3Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Czech RepublicPT05.Cryogenic-temperature electron microscopy imaging of extracellular vesicles shedding Naama Koifman1, Idan Biran1, Anat Aharon2, Benjamin Brenner3 and Yeshayahu Talmon1 Division of Chemical Engineering and the Russell Berrie Nanotechnology Institute (RBNI), Technion Israel Institute of Technology,.