o:36233 Three methods of isolating EVs from pleural effusion samples of patients with advanced lung adenocarcinoma-potential applications in clinical practice? en ABSTRACT Introduction: Pleural effusion (PE) occurs in 17-23% of lung cancer (LCa) patients and it contains extracellular vesicles (EVs) from cancer cells, representing “liquid biopsy” of LCa. Isolated PE-EVs could be used for LCa diagnosis or monitoring its progression/therapy. However, there is still no standard method for isolation of EVs from pleural fluid. The aim of this work is to compare PE-EVs isolation methods that could be employed both in research or clinical settings. Methods: PE samples diluted in PBS (1:1) from patients with advanced non-small cell lung cancer (NSCLC) were utilized. Three methods for isolating EVs were employed: an in-house spherical porous methacrylate-based copolymer coupled with VHH antibodies (chromatography method-CH), ultracentrifugation (UC), and the Norgen Plasma/Serum Exosome Purification and RNA Isolation Mini Kit (Commercial kit-CK). For each EVs isolation method, efficiency was monitored in terms of the amount of starting sample, time required for vesicle isolation, yield, quality of the obtained isolates and overall cost. Results: In terms of the amount of starting sample, the CH and CK have an advantage, allowing work with as little as 500μL, whereas UC required several milliliters of sample (in our case, due to the rotor type, 12mL minimum). Pleural effusion, extracellular vesicles, 1552100 978-86-905626-1-9 1552102 https://srbevs.rs/2nd-move-symposium/ 2025-06-05T06:07:17.553Z 44 yes 46 Miodrag Vuković Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia Lidija Filipović Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia person Nina Petrović Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia person Andrej Zečević Clinic for Pulmonology, University Clinical Center of Serbia, Belgrade, Serbia person Maja Kosanović University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor bioloških nauka person 0000-0002-1143-1805 Miljana Tanić Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia person Radmila Janković Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia person Tatjana Stanojković Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia person Milica Popović Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia person Aleksandra Korać Faculty of Biology, University of Belgrade, Belgrade, Serbia person Milena Čavić Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia person application/pdf 1608647 https://phaidrabg.bg.ac.rs/o:36233 no yes 1 70 92000004 11A19 11A1903 11A28 11A29 11A2904 11A02 2nd MOVE Symposium Abstract book, 8-11 October 2024, Belgrade, Serbia Serbian Society for Extracellular Vesicles, SrbEVs with Austrian Society for Extracellular Vesicles, ASEV Baltic Society for Extracellular Vesicles, BSEV Italian Society for Extracellular Vesicles, EVIta French Society for Extracellular Vesicles, FSEV Spanish Society for Extracellular Vesicles, GEIVEX German Society for Extracellular Vesicles, GSEV United Kingdom Society for Extracellular Vesicles, UKEV 2024