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High Sensitivity Phenotyping of Extracellular Vesicles by Customized, Single-Molecule Assays


May 11, 2023



Presented by Marina Cretich | Senior Researcher of the National Research Council of Italy at SCITEC-CNR

Key learning points

1. Extracellular Vesicles (EV) and exosomes are a promising avenue for liquid biopsy-based diagnostics and research but the sensitivity requirements make their use challenging
2. Customized assays have been developed for high sensitivity analysis of EV by Single Molecule Array technology (SiMoA) and compared with other digital platforms
3. SiMoA beads can be customized with probes other than antibodies and applied to EV phenotyping in various biological media


Extracellular Vesicles (EV) and exosomes, a subtype of EV, are tiny biogenic particles released by many types of cells in various bodily fluids such as blood, urine, and cerebrospinal fluid. As EVs contain and expose a variety of molecules, including proteins, lipids, and nucleic acids, they can serve as potential biomarkers for various diseases including cancer, neurological disorders, and infectious diseases. Although EV have the potential to be an excellent source of biomarkers in liquid biopsy, they face challenges in becoming a prominent choice due to their small size, low refractive index, inherent heterogeneity, and high sensitivity requirements to detect disease-specific sub-populations with low abundance. To meet these needs, innovative affinity-probes and digital detection techniques capable of achieving single-molecule sensitivity are required. In our recent work, we set customized assays for EV detection by Single Molecule Array technology (SiMoA) and compared the performance with Single Particle Interferometric Reflectance Imaging Sensing (SP-IRIS) and Super Resolution Microscopy. We will also discuss the potential to customize SiMoA beads with probes other than antibodies, providing examples of how this can be applied to EV phenotyping in various biological media.

About the Speaker

Marina Cretich is a Senior Researcher of the National Research Council of Italy at SCITEC-CNR in Milano, where she leads the projects on protein and peptide assays for diagnostics and biomarker discovery. Her research is focused on the development of new biomolecular methods for diagnosis and health monitoring, with a strong focus to system integration and technology development; ongoing projects include new analytical platforms for extracellular vesicles, point of care devices for infectious diseases, microarrays for allergy profiling and biomarker validation.She is currently coordinating the H2020 FET PROACTIVE project MARVEL (Evolving reversible iMmunocapture by membrane sensing peptides: towARds scalable extracellular VEsicLes isolation).She is member of the Board of Directors of EVIta (Italian Society for Extracellular Vesicles).