多肽识别导向的外泌体分离富集方法研究进展  

Research progress of peptide recognition-guided strategies for exosome isolation and enrichment

作　　者：徐坤 黄嫣嫣[1,2] 赵睿 XU Kun;HUANG Yanyan;ZHAO Rui(Beijing National Laboratory for Molecular Sciences,CAS Key Laboratory of Analytical Chemistry for Living Biosystems,CAS Research/Education Center for Excellence in Molecular Sciences,Institute of Chemistry,Chinese Academy of Sciences,Beijing 100190,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]北京分子科学国家研究中心,中国科学院活体分析化学重点实验室,中国科学院分子科学科教融合卓越创新中心,中国科学院化学研究所,北京100190 [2]中国科学院大学,北京100049

出　　处：《色谱》2025年第5期446-454,共9页Chinese Journal of Chromatography

基　　金：国家自然科学基金(22174145,22122411,22374152).

摘　　要：外泌体是由细胞分泌的双层膜囊泡,携带有丰富的生物分子,是细胞间信息交流的重要媒介,与多种疾病的发生发展密切相关。在复杂体系中开展外泌体分离富集研究,对于疾病诊断、预后检测和分子机制研究具有重要意义。但外泌体广泛存在于复杂生命体系中,尺寸小且分布范围宽,为其分离富集带来了困难与挑战。目前已经发展了基于物理性质的超速离心、体积排阻色谱、超滤和聚合物沉淀等方法。以亲和配体为识别单元,基于分子识别的亲和分离策略能够实现外泌体高选择性分离富集。在众多分子识别工具中,多肽凭借着可设计性强、构象自由灵活等优势,被应用于外泌体亲和分离之中。本文首先对目前外泌体分离富集方法进行了总结,之后重点介绍了以外泌体蛋白质和生物膜特性为靶标的亲和多肽设计与筛选原理及其在外泌体亲和分离富集中的应用。Exosomes are bilayered vesicles derived from living cells and bacteria that are loaded with abundant biomolecules,such as proteins and nucleic acids.As an important medium of remote cell communication,exosomes are closely related to the occurrence and development of a number of diseases,including those involving tumors and inflammation.The isolation and enrichment of exosomes in complex biosystems is greatly significant for the diagnosis,prognosis,and detection of diseases,as well as in molecular-mechanism research.However,exosomes are usually nanoscale size distribution and widely existed in complex biological samples,including blood,tissue fluids,and urine,which bring difficulties and challenges to the isolation and enrichment of exosomes.To address this issue,several methods based on the physical properties of exosomes have been developed.For example,exosomes can be obtained by ultracentrifugation at high centrifugal force based on density differences;they can also be isolated and enriched by size-exclusion chromatography and ultrafiltration based on size heterogeneity.Exosomes can also be separated in high yields but with low purities using commercial polymer-coprecipitation-based isolation kits.While the abovementioned methods can be used to isolate and enrich exosomes in a highly efficient manner,accurately distinguishing interfering particles,including protein aggregates and microvesicles,in biosystems is still difficult,resulting in the poor purity of exosome isolation and enrichment.Affinity ligands are widely used during the affinity isolation and enrichment of exosomes.Antibodies exhibit high selectivity and affinity;consequently exosomes can be captured highly selectively by exploiting specific antigen/antibody interactions.However,antibodies also have some limitations,including complex preparation procedures,high costs,and poor stability.Chemical affinity ligands,such as aptamers,peptides,and small molecules,are also widely used to isolate and enrich exosomes.As a kind of molecular recognition tool,p

关 键 词：外泌体 分子识别 多肽 分离富集 

分 类 号：O658[理学—分析化学]