基于微流控技术的外泌体分离分析及临床应用  

Exosome separation and analysis based on microfluidics technology and its clinical applications

作　　者：邢宇航 任香善[2] 李东浩[1] 刘璐 XING Yuhang;REN Xiangshan;LI Donghao;LIU Lu(Department of Chemistry,College of Science,Yanbian University,Key Laboratory of Natural Medicine Research in Changbaishan,Ministry of Education,Yanji 133000,China;Yanbian University Medical College,Yanji 133000,China)

机构地区：[1]延边大学理学院化学系,长白山天然药物研究教育部重点实验室,吉林延吉133000 [2]延边大学医学院,吉林延吉133000

出　　处：《色谱》2025年第5期455-471,共17页Chinese Journal of Chromatography

基　　金：国家自然科学基金项目(22304150,22376177);生物资源与功能分子学科创新引智基地(D18012);延边大学博士启动基金(ydbq202317).

摘　　要：外泌体是由细胞分泌的纳米级囊泡,其中富含脂质、蛋白质、核酸等多种生物功能分子。外泌体在生物体内的信息传递、疾病机理研究及诊断等方面扮演着类似“物联网、互联网”的关键角色。然而,受限于外泌体来源、尺寸、内含物以及功能的异质性,外泌体分离分析仍然面临诸多挑战。传统外泌体分离分析方法包括超速离心、尺寸排阻和免疫共沉淀等,但这些方法普遍存在产量低、纯度低等问题限制了其进一步的临床应用。微流控技术因其微型化、高通量、自动化和集成化等特点为复杂生物样本的高效分离分析提供了强有力的工具。基于此,本文系统总结了基于微流控技术的外泌体分离和分析方法。首先,讨论了基于外泌体的尺寸、电荷、表面官能团等物化性质结合微流控技术分离外泌体及外泌体亚群的分离方法;其次,分析了基于微流控平台的外泌体检测手段,包括外泌体群体分析方法和单个外泌体分析方法;最后,探讨了微流控技术在外泌体临床应用中的前景,并在疾病诊断和治疗方面进行了展望。微流控技术有望突破传统研究中的分离分析瓶颈,为癌症等重大疾病的精准诊断及治疗提供新技术、新平台。Exosomes are cell-secreted nanoscale vesicles 30-150 nm in size and encompass a diverse array of biomolecules,including lipids,proteins,and nucleic acids.Exosomes play pivotal roles during the intercellular exchange of materials and information,and are closely associated with the onset and progression of a variety of diseases.Therefore,comprehensively investigating exosomes is very important in terms of disease diagnosis and treatment.However,exosomes are genetically heterogeneous and are composed of different materials.Additionally,exosome-size and packing-specific-biomarker heterogeneities result in biofunction diversity.Moreover,isolating and analyzing exosomes is highly challenging owing to their small sizes and heterogeneities.Accordingly,effective separation methods and analytical techniques for highly specifically and efficiently identifying exosomes are urgently needed in order to better understand their functionalities.While separation and analysis is required to reveal exosome heterogeneity,the former is confronted by three primary challenges.Firstly,exosome heterogeneity(including heterogeneous marker expressions and size heterogeneity that results in heterogeneous functions)results in systems that are very difficult to separate.Secondly,the coexistence of non-vesicular contaminants(lipoprotein nanoparticles,soluble proteins,nucleic acids,etc.)and the complex matrix effects of body fluids also contribute to separation difficulties.Thirdly,enrichment is a highly challenging task owing to low exosome concentrations.Traditional methods,such as ultracentrifugation and size-exclusion chromatography,fall short in terms of their abilities to precisely separate and analyze exosomes.On the other hand,microfluidics has emerged as a robust tool for the efficient analysis of complex biological samples and is characterized by miniaturization,precise control,high throughput,automation,and integration.Firstly,the operability,integrability,and modifiability of a microfluidics system facilitate exosome separation and p

关 键 词：微流控 外泌体 分离分析 疾病诊断及治疗 

分 类 号：O658[理学—分析化学]