Valentina Marassia,b,c, Maria Serena Chiriacòc, Alessandro Romanod, Stefano Giordania, Anna Placcia, Barbara Rodaa,b, Andrea Zattonia,b, Pierluigi Reschigliana,b

aDepartment of Chemistry “G. Ciamician”, University of Bologna, Via Piero Gobetti 85, Bologna
bbyFlow srl, Viale Giuseppe Fanin 48, Bologna
cCNR NANOTEC - Institute of Nanotechnology, Via per Monteroni, 73100 Lecce, Italy
dInstitute of Experimental Neurology, San Raffaele Scientific Institute, 20132 Milan, Italy

Precision and personalized medicine aim to tailor treatments to the individual, taking into account each patient's unique biological profile. Liquid biopsy, by analysing biomarkers from biological fluids, can enhance these patient-centred approaches while providing a non-invasive alternative to traditional tissue biopsies. Extracellular vesicles (EVs), key mediators of intercellular communication, have emerged as powerful biomarkers due to their role in disease development. Despite their potential, the full understanding and application of EVs are hindered by significant limitations in current methods. Commonly used isolation techniques often demand large sample volumes and rely on time- and instrument-intensive processes. 

Characterization is also challenging due to EVs high heterogeneity and the need for sample pre-treatments, which can compromise vesicles integrity limiting their diagnostic and therapeutic potential. To address these limitations, we are working towards a streamlined approach that couples Flow Field-Flow Fractionation (4F) with on-chip detection, creating a benchtop-ready device capable of identifying arrays of biomarkers linked to EVs' subclasses size and function. Our goal is to minimize processing steps and analyse EVs in their native conditions, preserving their morphology and properties to enhance liquid biopsy effectiveness while reducing time and cost. Here we present preliminary findings on integrating dynamic cell culturing with our 4F platform and on-chip detection to streamline the isolation and analysis of EV-enriched fractions [1]. This approach aims to simplify processing, reduce analysis time, and minimize sensor interference, moving us closer to practical liquid biopsy applications.

Acknowledgements: Financed by the European Union – NextGeneration EU through the Italian Ministry of University and Research under PRIN 2022 RESOLVE, 202233FTW8 CUP B53D23013360006.

Keywords:  Flow Field-Flow Fractionation, Extracellular Vesicles, Liquid Biopsy

References

1. Marassi V, Giordani S, et al., Sensors, 23 (2023), pag. 9432. DOI: 10.3390/s23239432