色谱2025，Vol.43Issue(5)：455-471,17.DOI:10.3724/SP.J.1123.2024.10032
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
摘要
Abstract
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 piv-otal roles during the intercellular exchange of materials and information,and are closely associ-ated with the onset and progression of a variety of diseases.Therefore,comprehensively inves-tigating exosomes is very important in terms of disease diagnosis and treatment.However,exo-somes are genetically heterogeneous and are composed of different materials.Additionally,ex-osome-size and packing-specific-biomarker heterogeneities result in biofunction diversity.More-over,isolating and analyzing exosomes is highly challenging owing to their small sizes and het-erogeneities.Accordingly,effective separation methods and analytical techniques for highly spe-cifically 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 heterogene-ous 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 con-taminants(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 ul-tracentrifugation and size-exclusion chromatography,fall short in terms of their abilities to pre-cisely separate and analyze exosomes.On the other hand,microfluidics has emerged as a ro-bust tool for the efficient analysis of complex biological samples and is characterized by minia-turization,precise control,high throughput,automation,and integration.Firstly,the operabili-ty,integrability,and modifiability of a microfluidics system facilitate exosome separation and purification based on surface properties,size,charge,and polarity.Secondly,the use of a mi-crofluidics approach,with its high throughput,low reagent consumption,and multichannel ma-nipulability,greatly facilitates preparing exosomes and enhancing their concentrations.Thirdly,microfluidics ensures that diverse separation methods are compatible with downstream analysis techniques. Exosomes are highly heterogeneous;hence,they are classified by type and subpopulation(according to origin,size,molecular markers,functions,etc.).This paper first discusses mi-crofluidics techniques for separating exosomes and examines various separation strategies grounded in the physicochemical properties of exosomes.We then analyze exosome detection methodologies that use microfluidics platforms and encompass traditional group-exosome analy-sis techniques and novel single-exosome analysis approaches.Finally,we discuss future clinical applications of microfluidics technology in exosome research,particularly its potential for diag-nosing and treating diseases,thereby underscoring the applications value of microfluidics tech-nology in the realm of personalized and precision medicine.Furthermore,cutting-edge mi-crofluidics platforms offer novel perspectives for purifying and preparing EVs owing to precise fluid control,integration,miniaturization,and high-throughput characterization.EV popula-tions,subpopulations,and single vesicles can be purified based on their physicochemical prop-erties and microfluidics features.Comprehensive lab-on-a-chip methods are promising in terms of separating EVs based on traits,such as size,surface markers,and charge,and for obtaining highly pure EVs.Recycled EV samples can be prepared by controlling the high-throughput and multichannel capabilities of microfluidics approaches.The transition from bulk EV analysis to single-vesicle analysis provides opportunities to explore the heterogeneous nature of EVs,thereby augmenting their potential for disease diagnosis.关键词
微流控/外泌体/分离分析/疾病诊断及治疗Key words
microfluidics/exosomes/separation analysis/disease diagnosis and treatment分类
化学化工引用本文复制引用
邢宇航,任香善,李东浩,刘璐..基于微流控技术的外泌体分离分析及临床应用[J].色谱,2025,43(5):455-471,17.基金项目
国家自然科学基金项目(22304150,22376177) （22304150,22376177）
生物资源与功能分子学科创新引智基地(D18012) （D18012）
延边大学博士启动基金(ydbq202317).National Natural Science Foundation of China(Nos.22304150,22376177) （ydbq202317）
Biological Resources and Func-tional Molecular Science Innovation and Intelligence Base(No.D18012) （No.D18012）
Yanbian University Doctoral Foun-dation(No.ydbq202317). （No.ydbq202317）
