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A Fluorescence Assay for Exosome Detection Based on Bivalent Cholesterol Anchor Triggered Target Conversion and Enzyme-Free Signal Amplification

文献类型：期刊论文

作者Wang, Xiaokun5; Shang, Hezhen4; Ma, Cuiping5; Chen, Lingxin1,2,3
刊名ANALYTICAL CHEMISTRY
出版日期2021-06-22
卷号93期号:24页码:8493-8500
ISSN号0003-2700
DOI10.1021/acs.analchem.1c00796
通讯作者Ma, Cuiping(mcp169@163.com) ; Chen, Lingxin(lxchen@yic.ac.cn)
英文摘要Exosomes are emerging as one of the most promising biomarkers for early disease diagnosis and prognosis. The significant challenges facing the available methods include improving the detection specificity and sensitivity in complex biological samples. Herein, a fluorescence assay was established based on a combination of immunomagnetic separation and a two-step signal amplification strategy for direct isolation and subsequent detection of exosomes. First, immunomagnetic beads capture and enrich the exosomes via antibody-antigen reactions. Second, bivalent cholesterol (BC) anchors spontaneously insert into the lipid bilayer of bead-captured exosomes, forming a "one to many" amplification effect. The simultaneous recognition of the surface protein and the lipid bilayer structure of the exosome significantly eliminates the interference risk from free proteins. The detection of exosomes converts to the detection of BC-anchors. Finally, the sticky end of the BC-anchor acts as the initiator to trigger the enzyme-free DNA circuits for secondary signal amplification. Under the optimal conditions, highly sensitive and selective detection of exosomes was achieved ranging from 5.5 x 10(3) to 1.1 x 10(7) particles/mu L with a limit of detection of 1.29 x 10(3) particles/mu L. Moreover, this method allows the isolation and quantitative analysis of exosomes in several biological fluids with satisfactory recovery rates (92.25-106.8%). Thus, this approach provides a sensitive, anti-interference platform for isolating and detecting exosomes.
WOS关键词EXTRACELLULAR VESICLES ; CANCER EXOSOMES ; CELL ; SECRETION ; PROTEINS ; BIOGENESIS ; PROGRESS
资助项目National Nature Science Foundation of China[21904078] ; National Nature Science Foundation of China[21976209] ; Taishan Scholar Project Special Funding[ts20190962] ; Key Project of Natural Science Foundation of Shandong Province[ZR2020KH030]
WOS研究方向Chemistry
语种英语
WOS记录号WOS:000665642500016
资助机构National Nature Science Foundation of China ; Taishan Scholar Project Special Funding ; Key Project of Natural Science Foundation of Shandong Province
源URL[http://ir.yic.ac.cn/handle/133337/29464]  
专题烟台海岸带研究所_山东省海岸带环境工程技术研究中心
烟台海岸带研究所_中科院海岸带环境过程与生态修复重点实验室
通讯作者Ma, Cuiping; Chen, Lingxin
作者单位1.Binzhou Med Univ, Sch Pharm, Yantai 264003, Peoples R China
2.Qufu Normal Univ, Key Lab Life Organ Anal, Key Lab Pharmaceut Intermediates & Anal Nat Med, Coll Chem & Chem Engn, Qufu 273165, Shandong, Peoples R China
3.Chinese Acad Sci, Yantai Inst Coastal Zone Res, CAS Key Lab Coastal Environm Proc & Ecol Remediat, Shandong Key Lab Coastal Environm Proc, Yantai 264003, Peoples R China
4.Qingdao Chengyang Dist Peoples Hosp, Dept Hepatobiliary Surg, Qingdao 266109, Peoples R China
5.Qingdao Univ Sci & Technol, Shandong Prov Key Lab Biochem Engn, Coll Marine Sci & Biol Engn, Qingdao 266042, Peoples R China
推荐引用方式
GB/T 7714		 Wang, Xiaokun,Shang, Hezhen,Ma, Cuiping,et al. A Fluorescence Assay for Exosome Detection Based on Bivalent Cholesterol Anchor Triggered Target Conversion and Enzyme-Free Signal Amplification[J]. ANALYTICAL CHEMISTRY,2021,93(24):8493-8500.
APA Wang, Xiaokun,Shang, Hezhen,Ma, Cuiping,&Chen, Lingxin.(2021).A Fluorescence Assay for Exosome Detection Based on Bivalent Cholesterol Anchor Triggered Target Conversion and Enzyme-Free Signal Amplification.ANALYTICAL CHEMISTRY,93(24),8493-8500.
MLA Wang, Xiaokun,et al."A Fluorescence Assay for Exosome Detection Based on Bivalent Cholesterol Anchor Triggered Target Conversion and Enzyme-Free Signal Amplification".ANALYTICAL CHEMISTRY 93.24(2021):8493-8500.

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来源：烟台海岸带研究所

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