Comparison study of exosomes molecules driven from (NCI1975) NSCLC cell culture supernatant isolation and characterization techniques

Abstract

BackgroundLung cancer consider as one of leading cause of death around the world, normally the disease detection is difficult in early stages, Anew methods for diagnostic using non‐invasive way are developed using Nano‐molecules called exosomes. Exosomes are cell derived vesicles carrying different kind of molecules (proteins, DNA, RNA, microRNA) and also, displaying many proteins on their membrane surfaces. Those exosomes molecules are identical of parental cancer cells that induce them.MethodsIn this study, exosomes were isolated and characterized using multiple centrifugations, ultrafiltration, and chemical precipitation techniques. The resulted products were captured by two electro‐Microscopes (EM) methods for characterizing the morphology of exosomes as to differentiate exosomes from other extracellular vesicles Besides, the resulted products also further characterized by blotting the proteins extracted from exosomes against different monoclonal antibodies (CD81, CD63, CD9) that confirmed the existence of ligands attached to exosomes surfaces in western blot test.Results and Discussionthe exosomes isolated products were screened under Transmission Electron Microscopy (TEM). The size distributions of extracellular vesicles were significantly different from exosomes 160 nm P < 0.001). The size distributions of apoptotic vesicles (APV) 450nm and necrotic bodies, (NCB) 280 nm were significantly different from exosomes (P < 0.001). The same size band was observed in PVDF membranes of western blot test in all three methods using each monoclonal antibody. However, the filtration devices give higher protein yield and greater purity.In ConclusionsThe current results confirmed the accessible use of exosomes as biomarker to detect lung cancer cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.