Elaboration of a Bio-Plex platform for the characterization of Exosomes

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Extracellular vesicles (EVs) and exosomes are released by all cell types. A part of a heterogeneous family of vesicles, exosomes are vital in intercellular communication. They can be selectively taken up by neighboring or distant cells to deliver their cargo and contribute to the physiological or pathological functions of these cells. The field of EVs/Exosomes is expanding very rapidly, with an exponential increase in studies investigating their physiological and pathological functions. They can contribute to various chronic diseases including in ``Long COVID``symptoms or in Alzheimer's disease. Moreover, EVs/Exosomes could also be promising as novel nano-drug delivery tools.To date, the limitation in this field is the lack of the gold-standard method to harvest and to identify EVs/exosomes. They have to be characterized with several complementary technics for their size, morphology, charges..etc. However, considering the complexity of their biogenesis only the immune-detection of several specific markers could validate the presence of exosomes among other nanosized vesicles. Actually, the immunodetection of their markers following isolation is mostly limited to one or two analytes per sample and is time and sample-consuming. The originality of this proposal is to develop an ExoBio-Plex platform, designed to cover 14 EVs/Exosomal markers simultaneously with a very limited volume sample. Moreover, to better understand the role of EVs/exosomes in COVID-19 and in some chronic diseases, we will combine this platform to other Bio-Plex platforms such as the Bio-Plex MultiplexPro SARS-CoV-2 Assays, or Human Cytokine Screening Panel with 48-Plex, or with Alzheimer's related markers to develop respectively, the ExoCovid-19 Bio-Plex, the Exo-Cytokines-Bio-Plex, and the ExoAlz-Bio-Plex platforms. These platforms will be validated on different biofluids and cell culture media from different cell types. They will accelerate the fundamental and applied research on the role of EVs/Exosomes, will open new avenues for their application to other chronic diseases.