Elt University and by EFRO via the Interreg V Grensregio Vlaanderen Nederland project Trans Tech Diagnostics.LBP.Absolutely free flow electrophoresis allows preparation of EV fractions with higher recovery and SARS-CoV-2 Trimeric S Protein Proteins Recombinant Proteins purity prices Gerhard Weber1, Robert Wildgruber1, Simon Staubach2, Robin Dittrich3, Peter Horn3, Verena Boerger4 and Bernd Giebel2 FFE Service; 2Institute for Transfusion Medicine, University Hospital Essen, University of Duisburg-Essen, Essen, Germany; Division of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; 3Institute for Transfusion Medicine, University Hospital Essen, University of DuisburgEssen, Essen, German; 4Institute for Transfusion Medicine, University Hospital Essen, University Duisburg-Essen, Essen, GermanyIntroduction: Currently, it remains a challenge to prepare extracellular vesicles (EVs), in particular those from physique fluids, such as plasma, to high purity. Neither fractionation by density nor by size is sufficient to separate EVs from all contaminants e.g. high and low density lipoprotein (HDL/LDL) along with other contaminating components. For now, a timeconsuming combination of two techniques (density and size) is needed to enrich EVs to higher purities, on a regular basis resulting in low EV recoveries. Totally free Flow Electrophoresis is really a well-established preparative and micropreparative process to separate analytes with inherent difference of charge density and/or difference of pI-value. Approaches: Free of charge Flow Interval Zone Electrophoresis (FF-IZE), using media of distinctive pH-values, ranging from pH = eight to pH = four.eight delivers most appropriate protocols for the quantitative separation of amphoteric analytes,Thursday May perhaps 18,like proteins and peptides from non-amphoteric analytes like lipid vesicles, DNA and RNA. Outcomes: Inside our ongoing project we’ve got optimized FF-IZE-pH protocols for the purification and isolation of EVs as well DNA and RNA from cell culture supernatants and human plasma samples. Upon screening for EV-specific samples within a dot blot technique, EV-specific antigens are especially recovered in a chosen quantity of fractions. Presently, we characterize the identified fractions in more detail. For the enumeration of ready EVs we use the Nanoparticle Tracking Analysis (NTA). Additionally, the presence of EV markers and the absence of contaminants are analyzed by Western Blot. We document the appearance of isolated EVs by transmission electron microscopy and identify the miRNA profiles of your obtained fractions. Summary/Conclusion: The principle of FFE, the EV isolation approach and our ongoing final results will probably be presented.Funding Supported by the Polish National Centre for Study and Improvement Serpin B9 Proteins Recombinant Proteins STRATEGMED1/235773/19/NCBR/2016 “EXPLORE ME”.LBP.MicroRNA biogenesis and heterogeneous miRNA distribution in cancer EVs Nils J. Groenewegen, Catrin Lutz, Alba M. Losada, Monique A.J. van Eijndhoven and D. Michiel Pegtel Exosomes Research Group, Division of Pathology, VU University Medical Center, Amsterdam, The NetherlandsLBP.Visualization of extracellular vesicles derived from human bone marrow mesenchymal stem cells employing fluorescent and magnetic labels; in vitro and in vivo research Sylwia Koniusz1, Anna Andrzejewska1, Andrea Del Fattore2, Elbieta Karnas3, Malgorzata Frontczak-Baniewicz4, Hanna Kozlowska5, Maurizio Muraca6, Miroslaw Janowski7 and Barbara Lukomska1 NeuroRepair Department, Mossakowski Medical Investigation Centre, PAS, Warsaw, Poland; 2Multifactorial Illness and Complex Phenotype Research Region, Bambino GesChildren’.