基于电泳原理的外泌体纯化技术的研究

Exosomes are considered as important candidates for liquid biopsy of tumors. However, the main challenge in exosome research is the lack of simple, inexpensive, stable, and repeatable purification and enrichment methods. Developing an exosome separation technology with the advantages of high purity, easy operation, and low cost is highly desired. Lipoprotein is a major interference in the separation and purification of plasma exosomes. Therefore, removing the lipoprotein is the key technical difficulty to improve the purity of plasma exosome, since the size and density of lipoproteins are very close to exosomes. We speculate that under certain conditions, the electrophoretic rate of exosomes will be significantly different from that of lipoproteins, which can effectively separate exosomes from lipoproteins. Based on quantitative measurement and analysis of the electrical properties and size of exosome and lipoproteins, the aim of this study is to systematically explore the method of separating plasma exosomes and lipoproteins by gel electrophoresis. A combination of this technology with size-based exosome separation method will be helpful to further purify plasma exosomes and promote the research of cancer-related exosome biomarkers.

外泌体被认为是肿瘤液体活检中重要的研究对象。然而，外泌体研究中的主要挑战是缺乏简单、便宜且稳定和可重复的分离纯化和富集方法。开发一种能高效地从血浆中分离出高纯度外泌体，同时具备易操作、低成本等特点的外泌体分离技术是急需解决的问题。在血浆外泌体分离纯化过程中，血浆中的脂蛋白是重要的干扰物。因为脂蛋白的尺寸以及密度同外泌体非常相似，所以去除血浆脂蛋白对外泌体的污染是提高血浆外泌体纯度的关键技术难点。我们推测在某个特定条件下，外泌体电泳的速率将与其他脂蛋白电泳速率存在显著差异，从而可实现外泌体与脂蛋白的有效分离。本项目拟在定量检测分析外泌体和脂蛋白电性及尺寸的基础上，系统探索利用凝胶电泳技术分离血浆中外泌体和脂蛋白的方法。该技术与基于尺寸大小的外泌体分离技术相结合，将有助于深度纯化血浆外泌体，并促进癌症相关的外泌体标志物的研究。

细胞外囊泡（EV）被认为是肿瘤液体活检中重要的研究对象。然而，EV研究中的主要挑战是缺乏简单、便宜且稳定和可重复的分离纯化和富集方法。开发一种能高效地从血浆中分离出高纯度EV，同时具备易操作、低成本等特点的EV分离技术是急需解决的问题。在血浆EV分离纯化过程中，血浆中的脂蛋白是重要的干扰物。因为脂蛋白的尺寸以及密度同EV非常相似，所以去除血浆脂蛋白对EV的污染是提高血浆EV纯度的关键技术难点。我们的研究证明，EV电泳的速率与其他脂蛋白电泳速率存在显著差异，从而可实现EV与脂蛋白的有效分离。本项目在定量检测分析EV和脂蛋白电性及尺寸的基础上，系统探索利用凝胶电泳技术分离血浆中外EV和脂蛋白的方法。该技术与基于尺寸大小的EV分离技术相结合，帮助深度纯化血浆EV，并促进食管鳞状细胞癌相关的EV标志物的研究。