Tools for novel liquid biopsy approaches in Lung Cancer Diogo Fortunatoa, Laura Bianciardib, Gaia Papinic, Mattia Criscuolic, Davide Zoccod and Natasa Zarovnib Exosomics/University of Siena; bExosomics; cExosomics Siena, University of Siena; dExosomics SienaaSide scatter module for enhanced detection of Extracellular Vesicles by flow cytometry. Tina Van Den Broecka, Ludovic Monheimb, Ihor Berezhnyya, Oleg Guryeva, Tatyana Chernenkoa, Marybeth Sharkeya and Geoffrey OsborneaaIntroduction: Mounting clinical evidence suggests that liquid biopsy may revolutionize the way cancer individuals are presently managed. Inside this context, our study aims to assess and reinforce exclusive and complementary positive aspects of EV/exosome-based approaches, by means of identification and quantitative detection of non-small cell lung cancer (NSCLC) EV biomarkers. Current technologies and methods for exosome isolation from complicated biological samples (i.e. plasma), have shown to become unreliable. There’s a really need to substantially increase them to allow multiparameter EV evaluation. Therefore, moreover to EV-biomarker discovery, we’re testing plasma processing and preanalytical tools, devices and optimized immunoaffinity protocols that tackle fundamental obstacles, like complicated matrix effects. Our goal is always to present an EV immunocapture approach with sufficient sensitivity, specificity and robustness for clinical grade diagnostic applications. Strategies: Size-based vs. immunocapture strategies for exosome isolation. Enzymatic and immunological Fc-gamma Receptor I/CD64 Proteins Molecular Weight assays for plasma pre-clearing; Flow cytometry, ELISA, nanoparticle tracking evaluation, Western Blot, SPR and ddPCR for antibody and exosome characterization. Final
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