{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T12:48:09Z","timestamp":1785847689061,"version":"3.56.0"},"reference-count":35,"publisher":"American Chemical Society (ACS)","issue":"30","license":[{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/na","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22074032"],"award-info":[{"award-number":["22074032"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22574047"],"award-info":[{"award-number":["22574047"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"Open Funding of Key Laboratory of Monitoring for Heavy Metal Pollutants, Ministry of Ecology and Environment","doi-asserted-by":"publisher","award":["KLMHM202504"],"award-info":[{"award-number":["KLMHM202504"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Exosomes, rich in diverse bioactive molecules, have emerged as a promising \u201cliquid biopsy\u201d approach for early cancer screening. However, relying solely on a single biomarker type can easily lead to missed diagnoses or misjudgments. Therefore, there is an urgent need to develop multiple biomarkers detection strategies to facilitate precise diagnosis. Herein, a microfluidic chip integrating two signals (ALP-driven chemiluminescence and nanobeacons-based fluorescence) into a single device was developed using breast cancer exosomes as a model. A simple sample injection enabled capture and quantitative detection of exosomal surface proteins (CD63, EpCAM, HER2) and internal miRNAs (miRNA-21, miRNA-375). After capture, protein levels were determined by driving magnetic beads and capturing images with a smartphone. Prefabricated nanobeacons were then directly introduced into the chip, enabling miRNA signal detection without RNA extraction. This five-marker biomarker panel achieved a diagnostic accuracy of 99.6% in breast cancer identification, representing an improvement of at least 4.6% over single-marker approaches and an 8.6% increase over any individual biomarker. This multitarget exosome analysis platform may provide a highly sensitive, low-cost, and easily scalable novel tool for breast cancer early diagnosis and treatment efficacy assessment.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c02416","type":"journal-article","created":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T11:23:46Z","timestamp":1784805826000},"page":"22291-22299","source":"Crossref","is-referenced-by-count":0,"title":["Multiplexed\nDetection of Breast Cancer-Specific miRNA\nand Protein Biomarkers on Exosomes Using a Microfluidic Platform"],"prefix":"10.1021","volume":"98","author":[{"given":"Hongqiang","family":"Wang","sequence":"first","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Liu","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanan","family":"Luan","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Du","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Wang","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Yang","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8122-7140","authenticated-orcid":true,"given":"Xiaohai","family":"Yang","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7337-0999","authenticated-orcid":true,"given":"Qing","family":"Wang","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9390-4938","authenticated-orcid":true,"given":"Kemin","family":"Wang","sequence":"additional","affiliation":[{"name":"Hunan University , , ,","place":["Changsha, China, 410082"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,23]]},"reference":[{"key":"2026080408082593400_cit1","doi-asserted-by":"publisher","first-page":"116060","DOI":"10.1016\/j.bios.2024.116060","article-title":"An Electrochemical Biosensor Designed\nwith Entropy-Driven Autocatalytic DNA Circuits for Sensitive Detection\nof Ovarian Cancer-Derived Exosomes","volume":"250","author":"Deng","year":"2024","journal-title":"Biosens.\nBioelectron."},{"issue":"6478","key":"2026080408082593400_cit2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aau6977","article-title":"The Biology, Function,\nand Biomedical Applications\nof Exosomes","volume":"367","author":"Kalluri","year":"2020","journal-title":"Science"},{"issue":"19","key":"2026080408082593400_cit3","doi-asserted-by":"publisher","first-page":"12035","DOI":"10.1021\/acs.chemrev.0c01140","article-title":"Aptamer-Based Detection of Circulating\nTargets for Precision Medicine","volume":"121","author":"Wu","year":"2021","journal-title":"Chem. Rev."},{"issue":"6","key":"2026080408082593400_cit4","doi-asserted-by":"publisher","first-page":"5017","DOI":"10.1021\/acsnano.3c11147","article-title":"Single-Cell\nLiquid\nBiopsy of Lung Cancer: Ultra-Simplified Efficient Enrichment of Circulating\nTumor Cells and Hand-Held Fluorometer Portable Testing","volume":"18","author":"He","year":"2024","journal-title":"ACS Nano"},{"issue":"8","key":"2026080408082593400_cit5","doi-asserted-by":"publisher","first-page":"2409036","DOI":"10.1002\/smll.202409036","article-title":"An Advanced\nMultivalent Ligand-Decorated\nMicrosphere Enrichment System Efficiently Captures Circulating Tumor\nCells In Vivo","volume":"21","author":"Wang","year":"2025","journal-title":"Small"},{"issue":"1","key":"2026080408082593400_cit6","doi-asserted-by":"publisher","first-page":"56","DOI":"10.1186\/s12943-022-01509-9","article-title":"Exosomes as\na New Frontier of Cancer\nLiquid Biopsy","volume":"21","author":"Yu","year":"2022","journal-title":"Mol. Cancer"},{"issue":"1","key":"2026080408082593400_cit7","doi-asserted-by":"publisher","first-page":"1271","DOI":"10.1021\/acsnano.4c13557","article-title":"Concurrent Detection\nof Protein and miRNA at the Single Extracellular Vesicle Level Using\na Digital Dual CRISPR-Cas Assay","volume":"19","author":"Xu","year":"2025","journal-title":"ACS Nano"},{"issue":"2","key":"2026080408082593400_cit8","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12404","article-title":"Minimal Information\nfor Studies of Extracellular Vesicles (MISEV2023): From Basic to Advanced\nApproaches","volume":"13","author":"Welsh","year":"2024","journal-title":"J. Extracell. Vesicles"},{"issue":"12","key":"2026080408082593400_cit9","doi-asserted-by":"publisher","first-page":"10737","DOI":"10.1021\/jacs.5c01973","article-title":"Profiling\nNascent Tumor\nExtracellular Vesicles via Metabolic Timestamping and Aptamer-Driven\nSpecific Click Chemistry","volume":"147","author":"Wu","year":"2025","journal-title":"J. Am. Chem. Soc."},{"issue":"24","key":"2026080408082593400_cit10","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202303121","article-title":"Selective in Situ Analysis of Mature microRNAs in Extracellular\nVesicles Using a DNA Cage-Based Thermophoretic Assay","volume":"62","author":"Zhao","year":"2023","journal-title":"Angew. Chem., Int. Ed."},{"issue":"19","key":"2026080408082593400_cit11","doi-asserted-by":"publisher","first-page":"18630","DOI":"10.1021\/acsnano.5c02864","article-title":"Accurate Cancer\nDiagnosis and Treatment Monitoring through Multiplexed Profiling of\nProtein Markers on Small Extracellular Vesicles","volume":"19","author":"Ren","year":"2025","journal-title":"ACS Nano"},{"issue":"38","key":"2026080408082593400_cit12","doi-asserted-by":"publisher","first-page":"15177","DOI":"10.1021\/acs.analchem.4c02062","article-title":"Fast Isolation and\nSensitive Multicolor Visual Detection of Small Extracellular Vesicles\nby Multifunctional Polydopamine Nanospheres","volume":"96","author":"Zhang","year":"2024","journal-title":"Anal. Chem."},{"issue":"7","key":"2026080408082593400_cit13","doi-asserted-by":"publisher","first-page":"5404","DOI":"10.1021\/acs.analchem.0c00106","article-title":"Magneto-Mediated Electrochemical\nSensor for Simultaneous Analysis of Breast Cancer Exosomal Proteins","volume":"92","author":"An","year":"2020","journal-title":"Anal. Chem."},{"key":"2026080408082593400_cit14","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1016\/j.bios.2019.04.013","article-title":"Direct Quantification\nof Cancerous\nExosomes via Surface Plasmon Resonance with Dual Gold Nanoparticle-Assisted\nSignal Amplification","volume":"135","author":"Wang","year":"2019","journal-title":"Biosens. Bioelectron."},{"issue":"5","key":"2026080408082593400_cit15","doi-asserted-by":"publisher","first-page":"5526","DOI":"10.1021\/acsnano.4c14209","article-title":"Digital\nProfiling of Tumor Extracellular Vesicle-Associated RNAs Directly\nfrom Unprocessed Blood Plasma","volume":"19","author":"Clarissa","year":"2025","journal-title":"ACS Nano"},{"issue":"42","key":"2026080408082593400_cit16","doi-asserted-by":"publisher","first-page":"15331","DOI":"10.1021\/acs.nanolett.5c04034","article-title":"Integrated Chemical Array and SERS Profiling of Plasma Small Extracellular\nVesicles for Breast Cancer Diagnosis","volume":"25","author":"Feng","year":"2025","journal-title":"Nano Lett."},{"issue":"17","key":"2026080408082593400_cit17","doi-asserted-by":"publisher","first-page":"9212","DOI":"10.1021\/acs.analchem.4c06417","article-title":"Enrichment\nand Detection of HER2-Expressing Extracellular Vesicles\nBased on DNA Tetrahedral Nanostructures: A New Strategy for Liquid\nBiopsy in Breast Cancer","volume":"97","author":"Xu","year":"2025","journal-title":"Anal. Chem."},{"issue":"13","key":"2026080408082593400_cit18","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abm3088","article-title":"Sensitive\nDetection of Multiple Blood Biomarkers via\nImmunomagnetic Exosomal PCR for the Diagnosis of Alzheimer\u2019s\nDisease","volume":"10","author":"Hu","year":"2024","journal-title":"Sci. Adv."},{"issue":"27","key":"2026080408082593400_cit19","doi-asserted-by":"publisher","first-page":"2301930","DOI":"10.1002\/advs.202370180","article-title":"Inaugurating\nHigh-Throughput Profiling of Extracellular\nVesicles for Earlier Ovarian Cancer Detection","volume":"10","author":"Jo","year":"2023","journal-title":"Adv. Sci."},{"issue":"4","key":"2026080408082593400_cit20","doi-asserted-by":"publisher","DOI":"10.1002\/ange.202314262","article-title":"An Aptamer-Based Nanoflow\nCytometry Method for the Molecular Detection and Classification of\nOvarian Cancers through Profiling of Tumor Markers on Small Extracellular\nVesicles","volume":"136","author":"Li","year":"2024","journal-title":"Angew. Chem., Int. Ed."},{"issue":"48","key":"2026080408082593400_cit21","doi-asserted-by":"publisher","first-page":"2004492","DOI":"10.1002\/smll.202004492","article-title":"Accurate Cancer\nDiagnosis and Stage Monitoring Enabled by Comprehensive Profiling\nof Different Types of Exosomal Biomarkers: Surface Proteins and miRNAs","volume":"16","author":"Zhou","year":"2020","journal-title":"Small"},{"issue":"51","key":"2026080408082593400_cit22","doi-asserted-by":"publisher","first-page":"18034","DOI":"10.1021\/acs.analchem.2c04408","article-title":"Noninvasive\nMultiplexed Analysis of Bladder Cancer-Derived Urine Exosomes via\nJanus Magnetic Microspheres","volume":"94","author":"Wei","year":"2022","journal-title":"Anal. Chem."},{"issue":"23","key":"2026080408082593400_cit23","doi-asserted-by":"publisher","first-page":"23584","DOI":"10.1021\/acsnano.3c06604","article-title":"Nanoprojectile\nSecondary Ion Mass Spectrometry Enables Multiplexed Analysis of Individual\nHepatic Extracellular Vesicles","volume":"17","author":"Lee","year":"2023","journal-title":"ACS Nano"},{"issue":"3","key":"2026080408082593400_cit24","doi-asserted-by":"publisher","first-page":"1555","DOI":"10.1021\/acssensors.3c02789","article-title":"Deep Learning Promotes Profiling of Multiple miRNAs in Single Extracellular\nVesicles for Cancer Diagnosis","volume":"9","author":"Zhang","year":"2024","journal-title":"ACS Sens."},{"issue":"1","key":"2026080408082593400_cit25","doi-asserted-by":"publisher","first-page":"4278","DOI":"10.1038\/s41467-023-39844-0","article-title":"Multiplexed\nRNA Profiling by Regenerative Catalysis Enables Blood-Based Subtyping\nof Brain Tumors","volume":"14","author":"Zhang","year":"2023","journal-title":"Nat. Commun."},{"key":"2026080408082593400_cit26","doi-asserted-by":"publisher","first-page":"115592","DOI":"10.1016\/j.bios.2023.115592","article-title":"An Immuno-Magnetophoresis-Based\nMicrofluidic Chip to Isolate and Detect HER2-Positive Cancer-Derived\nExosomes via Multiple Separation","volume":"239","author":"Mun","year":"2023","journal-title":"Biosens. Bioelectron."},{"issue":"17","key":"2026080408082593400_cit27","doi-asserted-by":"publisher","first-page":"6618","DOI":"10.1021\/acs.analchem.3c05858","article-title":"Codetection of Proteins\nand RNAs on Extracellular Vesicles for Pancreatic Cancer Early Diagnosis","volume":"96","author":"He","year":"2024","journal-title":"Anal. Chem."},{"issue":"11","key":"2026080408082593400_cit28","doi-asserted-by":"publisher","first-page":"2306373","DOI":"10.1002\/advs.202306373","article-title":"Extracellular Vesicular Analysis of Glypican 1 mRNA\nand Protein for Pancreatic Cancer Diagnosis and Prognosis","volume":"11","author":"Li","year":"2024","journal-title":"Adv. Sci."},{"issue":"12","key":"2026080408082593400_cit29","doi-asserted-by":"publisher","first-page":"9256","DOI":"10.1021\/acssensors.5c00605","article-title":"Visual Quantitative\nDetection of Tumor-Derived Exosomes Based on Catalyzed Decomposition\nof H2O2 Using a Portable Microchannel Device","volume":"10","author":"Guan","year":"2025","journal-title":"ACS Sens."},{"key":"2026080408082593400_cit30","doi-asserted-by":"publisher","first-page":"113826","DOI":"10.1016\/j.bios.2021.113826","article-title":"The Activity of\nAlkaline Phosphatase in Breast Cancer\nExosomes Simplifies the Biosensing Design","volume":"198","author":"Moura","year":"2022","journal-title":"Biosens.\nBioelectron."},{"issue":"11","key":"2026080408082593400_cit31","doi-asserted-by":"publisher","first-page":"4996","DOI":"10.1021\/jacs.9b13960","article-title":"Thermophoretic Detection\nof Exosomal microRNAs by Nanoflares","volume":"142","author":"Zhao","year":"2020","journal-title":"J. Am.\nChem. Soc."},{"key":"2026080408082593400_cit32","doi-asserted-by":"publisher","first-page":"339995","DOI":"10.1016\/j.aca.2022.339995","article-title":"Polymer-Assisted Au@PDA Nanoparticles\nLyophilized Powder with High Stability and Low Adsorption and Its\nApplication in Colorimetric Biosensing","volume":"1220","author":"Wang","year":"2022","journal-title":"Anal.\nChim. Acta"},{"issue":"3","key":"2026080408082593400_cit33","doi-asserted-by":"publisher","first-page":"1709","DOI":"10.1021\/acs.analchem.0c04308","article-title":"Precise Capture and Direct Quantification\nof Tumor Exosomes via a Highly Efficient Dual-Aptamer\nRecognition-Assisted Ratiometric Immobilization-Free Electrochemical\nStrategy","volume":"93","author":"Yang","year":"2021","journal-title":"Anal. Chem."},{"issue":"30","key":"2026080408082593400_cit34","doi-asserted-by":"publisher","first-page":"13475","DOI":"10.1021\/jacs.2c00119","article-title":"Molecular Characterization\nof Exosomes for Subtype-Based Diagnosis of Breast Cancer","volume":"144","author":"Cao","year":"2022","journal-title":"J. Am. Chem. Soc."},{"issue":"50","key":"2026080408082593400_cit35","doi-asserted-by":"publisher","first-page":"27959","DOI":"10.1021\/acs.analchem.5c05657","article-title":"Label-Free\nProbes and Magneto-Thermal Imprinting Enable Rapid Multiplexed Exosomal\nProfiling for Breast Cancer Classification","volume":"97","author":"Zhao","year":"2025","journal-title":"Anal. Chem."}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.6c02416","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/30\/22291\/66221196\/acs.analchem.6c02416.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/30\/22291\/66221196\/acs.analchem.6c02416.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T12:20:35Z","timestamp":1785846035000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/98\/30\/22291\/5230392\/Multiplexed-Detection-of-Breast-Cancer-Specific"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,23]]},"references-count":35,"journal-issue":{"issue":"30","published-online":{"date-parts":[[2026,7,23]]},"published-print":{"date-parts":[[2026,8,4]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.6c02416","relation":{},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,23]]}}}