Raizada Geetika, Namasivayam Balasubramaniam, Obeid Sameh, Brunel Benjamin, Boireau Wilfrid, Lesniewska Eric, Elie-Caille Celine
Université de Franche-Comté, CNRS UMR-6174, FEMTO-ST Institute.
Lille Neuroscience & Cognition research centre.
J Vis Exp. 2023 Mar 17(193). doi: 10.3791/64210.
Extracellular vesicles (EVs) are membrane-derived, tiny vesicles produced by all cells that range from 50 to several hundreds of nanometers in diameter and are used as a means of intercellular communication. They are emerging as promising diagnostic and therapeutic tools for a variety of diseases. There are two main biogenesis processes used by cells to produce EVs with differences in size, composition, and content. Due to their high complexity in size, composition, and cell origin, their characterization requires a combination of analytical techniques. This project involves the development of a new generation of multiparametric analytical platforms with increased throughput for the characterization of subpopulations of EVs. To achieve this goal, the work starts from the nanobioanalytical platform (NBA) established by the group, which allows an original investigation of EVs based on a combination of multiplexed biosensing methods with metrological and morphomechanical analyses by atomic force microscopy (AFM) of vesicular targets trapped on a microarray biochip. The objective was to complete this EV investigation with a phenotypic and molecular analysis by Raman spectroscopy. These developments enable the proposal of a multimodal and easy-to-use analytical solution for the discrimination of EV subsets in biological fluids with clinical potential.
细胞外囊泡(EVs)是由所有细胞产生的膜衍生微小囊泡,直径范围从50到数百纳米,用作细胞间通讯的一种方式。它们正成为多种疾病有前景的诊断和治疗工具。细胞产生EVs主要有两种生物发生过程,其大小、组成和内容物存在差异。由于其在大小、组成和细胞来源方面高度复杂,对其进行表征需要多种分析技术的结合。该项目涉及开发新一代多参数分析平台,以提高对EVs亚群表征的通量。为实现这一目标,工作从该团队建立的纳米生物分析平台(NBA)开始,该平台允许基于多重生物传感方法与通过原子力显微镜(AFM)对捕获在微阵列生物芯片上的囊泡靶标进行计量和形态力学分析相结合,对EVs进行原创性研究。目标是通过拉曼光谱进行表型和分子分析来完成对EVs的研究。这些进展使得能够提出一种多模态且易于使用的分析解决方案,用于区分具有临床潜力的生物流体中的EV亚群。