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

细胞外囊泡（EVs）和外泌体由所有细胞类型释放。作为异质囊泡家族的一部分，外泌体在细胞间通讯中起着至关重要的作用。它们可以被邻近或远处的细胞选择性地吸收，以运送它们的货物，并有助于这些细胞的生理或病理功能。ev /外泌体的研究领域正在迅速扩大，对其生理和病理功能的研究呈指数级增长。它们可能导致“长冠状病毒”症状或阿尔茨海默病等各种慢性疾病。此外，ev /外泌体也有望成为新型纳米药物输送工具。迄今为止，该领域的局限性在于缺乏采集和鉴定ev /外泌体的金标准方法。它们必须用几种互补的技术来表征它们的尺寸、形态、电荷等。然而，考虑到其生物发生的复杂性，只有几种特定标记的免疫检测才能验证外泌体在其他纳米囊泡中的存在。实际上，分离后对其标记物的免疫检测大多局限于每个样品一到两个分析物，并且耗时耗样。该提案的独创性在于开发ExoBio-Plex平台，旨在以非常有限的体积样品同时覆盖14个ev /外泌体标记物。此外，为了更好地了解ev /外泌体在COVID-19和一些慢性疾病中的作用，我们将把该平台与其他Bio-Plex平台（如Bio-Plex MultiplexPro SARS-CoV-2检测、Human Cytokine Screening Panel with 48-Plex或与阿尔茨海默氏症相关标志物）结合起来，分别开发ExoCovid-19 Bio-Plex、Exo-Cytokines-Bio-Plex和ExoAlz-Bio-Plex平台。这些平台将在来自不同细胞类型的不同生物流体和细胞培养基上进行验证。它们将加速ev /外泌体作用的基础和应用研究，为其在其他慢性疾病中的应用开辟新的途径。

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.