{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T09:06:52Z","timestamp":1789376812400,"version":"build-2803163510"},"reference-count":107,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,13]],"date-time":"2026-09-13T00:00:00Z","timestamp":1789257600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Major Special Program of Gansu Province","award":["26ZDFA003"],"award-info":[{"award-number":["26ZDFA003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Extracellular vesicles (EVs) have dominated research on intercellular communication for decades. However, the discovery of exomeres in 2018 revealed that conventional exosome preparations contain large quantities of long-overlooked non-membranous nanoparticles. Additional non-vesicular extracellular nanoparticles (NVEPs), including supermeres and SECmeres, have since been reported. Most of these particles are smaller than 50 nm in diameter, yet they carry protein and RNA cargo that was previously thought to be delivered exclusively by exosomes. Consequently, certain functions once attributed to exosomes now require reassignment. This review summarizes the classification, cargo composition, and structural features of NVEPs. It then compares the respective limitations of differential ultracentrifugation, density gradient centrifugation, size-exclusion chromatography, and flow field-flow fractionation for NVEP isolation, together with strategies for their combined use. Furthermore, the functional mechanisms that distinguish NVEPs from vesicular EVs are discussed at three levels: the extracellular stability of non-vesicular extracellular RNA (nv-RNA), cellular uptake pathways, and downstream effects. Current evidence indicates that NVEPs are enriched for disease-associated cargo in cancer and neurodegenerative diseases and show potential for liquid biopsy and drug delivery. Nevertheless, ambiguous definitional boundaries, the absence of reference materials, and insufficient in vivo evidence jointly hinder quantification and clinical translation in this field. Because these limitations arise largely from the absence of unified technical criteria, establishing internationally recognized standards for isolation and characterization is a prerequisite for the maturation of NVEP research.<\/jats:p>","DOI":"10.3390\/biom16091330","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T08:10:23Z","timestamp":1789373423000},"page":"1330","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Beyond Exosomes: Biological Properties, Isolation Challenges, and Functional Redefinition of Non-Vesicular Extracellular Nanoparticles (NVEPs)"],"prefix":"10.3390","volume":"16","author":[{"given":"Hengxiang","family":"Liu","sequence":"first","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zishuai","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hai","family":"He","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaxue","family":"Yuan","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyong","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heqiang","family":"Yang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sijing","family":"Yang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4859-6926","authenticated-orcid":false,"given":"Zifan","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, School of Bioengineering, Northwest Minzu University, Lanzhou 730030, China"},{"name":"Gansu Animal Cell Technology Innovation Center, Northwest Minzu University, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiqiang","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Oral Diseases of Gansu Province, Key Laboratory of Stomatology of State Ethnic Affairs Commission, School of Stomatology, Northwest Minzu University, No. 1 Xibeixincun Baiyin Road, Chengguan District, Lanzhou 730030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e12386","DOI":"10.1002\/jev2.12386","article-title":"Exploring the role of polymers to overcome ongoing challenges in the field of extracellular vesicles","volume":"12","author":"Simon","year":"2023","journal-title":"J. Extracell. Vesicles"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/bs.acc.2021.07.008","article-title":"Extracellular vesicles in respiratory disease","volume":"108","author":"Carnino","year":"2022","journal-title":"Adv. Clin. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.cytogfr.2023.09.006","article-title":"Extracellular vesicles in cardiovascular diseases: From pathophysiology to diagnosis and therapy","volume":"74","author":"Du","year":"2023","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"105392","DOI":"10.1016\/j.critrevonc.2026.105392","article-title":"Exosomes and extracellular vesicles in gastric cancer immune escape","volume":"224","author":"Mazzone","year":"2026","journal-title":"Crit. Rev. Oncol.\/Hematol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"216960","DOI":"10.1016\/j.canlet.2024.216960","article-title":"Revisiting the advances and challenges in the clinical applications of extracellular vesicles in cancer","volume":"593","author":"He","year":"2024","journal-title":"Cancer Lett."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1038\/s41565-024-01774-3","article-title":"Harnessing extracellular vesicle heterogeneity for diagnostic and therapeutic applications","volume":"20","author":"Carney","year":"2025","journal-title":"Nat. Nanotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"11619","DOI":"10.1021\/acsnano.2c04337","article-title":"Future of Digital Assays to Resolve Clinical Heterogeneity of Single Extracellular Vesicles","volume":"16","author":"Morales","year":"2022","journal-title":"ACS Nano"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e70005","DOI":"10.1002\/jev2.70005","article-title":"Extracellular vesicles, RNA sequencing, and bioinformatic analyses: Challenges, solutions, and recommendations","volume":"13","author":"Miceli","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1186\/s12951-025-03354-6","article-title":"Extracellular particles: Emerging insights into central nervous system diseases","volume":"23","author":"Chen","year":"2025","journal-title":"J. Nanobiotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e1835","DOI":"10.1002\/wnan.1835","article-title":"Isolation and characterization of extracellular vesicles and future directions in diagnosis and therapy","volume":"15","author":"Rossi","year":"2023","journal-title":"Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol."},{"key":"ref_11","first-page":"322","article-title":"Exomeres and supermeres: Current advances and perspectives","volume":"50","author":"Yu","year":"2025","journal-title":"Bioact. Mater."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1016\/j.tcb.2023.01.002","article-title":"Extracellular vesicles and nanoparticles: Emerging complexities","volume":"33","author":"Jeppesen","year":"2023","journal-title":"Trends Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1038\/s41556-018-0040-4","article-title":"Identification of distinct nanoparticles and subsets of extracellular vesicles by asymmetric flow field-flow fractionation","volume":"20","author":"Zhang","year":"2018","journal-title":"Nat. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1462","DOI":"10.1038\/s41596-023-00811-0","article-title":"Comprehensive isolation of extracellular vesicles and nanoparticles","volume":"18","author":"Zhang","year":"2023","journal-title":"Nat. Protoc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1038\/s41556-021-00805-8","article-title":"Supermeres are functional extracellular nanoparticles replete with disease biomarkers and therapeutic targets","volume":"23","author":"Zhang","year":"2021","journal-title":"Nat. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5453","DOI":"10.1038\/s41467-026-74541-8","article-title":"SECmeres outperform extracellular vesicles as potential blood RNA biomarkers for Alzheimer\u2019s disease","volume":"17","author":"Tichkule","year":"2026","journal-title":"Nat. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"108774","DOI":"10.1016\/j.pharmthera.2024.108774","article-title":"Non-vesicular extracellular RNA: A potential drug target to intervene cell-cell communication","volume":"266","author":"Tomita","year":"2025","journal-title":"Pharmacol. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"15554","DOI":"10.1039\/D6NR01765G","article-title":"Tangential flow filtration for isolating exomeres and other nanoscale extracellular particles","volume":"18","author":"Scarborough","year":"2026","journal-title":"Nanoscale"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1039\/D4NH00320A","article-title":"Extracellular vesicles of different cellular origin feature distinct biomolecular corona dynamics","volume":"10","author":"Zendrini","year":"2025","journal-title":"Nanoscale Horiz."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e70169","DOI":"10.1002\/jev2.70169","article-title":"Exploring the Spatial Limits of Extracellular Vesicles-Mediated Intercellular Communication","volume":"14","author":"Colombo","year":"2025","journal-title":"J. Extracell. Vesicles"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e12140","DOI":"10.1002\/jev2.12140","article-title":"Formation of a protein corona on the surface of extracellular vesicles in blood plasma","volume":"10","author":"Visnovitz","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e12207","DOI":"10.1002\/jev2.12207","article-title":"A functional corona around extracellular vesicles enhances angiogenesis, skin regeneration and immunomodulation","volume":"11","author":"Wolf","year":"2022","journal-title":"J. Extracell. Vesicles"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e12399","DOI":"10.1002\/jev2.12399","article-title":"Uptake of extracellular vesicles into immune cells is enhanced by the protein corona","volume":"12","author":"Dietz","year":"2023","journal-title":"J. Extracell. Vesicles"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1038\/s41565-023-01585-y","article-title":"Cellular uptake and in vivo distribution of mesenchymal-stem-cell-derived extracellular vesicles are protein corona dependent","volume":"19","author":"Faruqu","year":"2024","journal-title":"Nat. Nanotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e21913","DOI":"10.1002\/advs.202521913","article-title":"Beyond Extracellular Vesicle (EV) Hype: Practical Solutions and Remaining Hurdles in EV Research, Manufacturing, and Clinical Translation","volume":"13","author":"Lundy","year":"2026","journal-title":"Adv. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e70209","DOI":"10.1002\/jev2.70209","article-title":"Unveiling Heterogeneity: Innovations and Challenges in Single-Vesicle Analysis for Clinical Translation","volume":"14","author":"Zhang","year":"2025","journal-title":"J. Extracell. Vesicles"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"189210","DOI":"10.1016\/j.bbcan.2024.189210","article-title":"The emerging role of extracellular vesicles and particles in prostate cancer diagnosis, and risk stratification","volume":"1879","author":"Wang","year":"2024","journal-title":"Biochim. Biophys. Acta Rev. Cancer"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e70168","DOI":"10.1002\/jev2.70168","article-title":"Heterogeneity of Extracellular Vesicles and Non-Vesicular Nanoparticles in Glioblastoma","volume":"14","author":"Xu","year":"2025","journal-title":"J. Extracell. Vesicles"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"116287","DOI":"10.1016\/j.celrep.2025.116287","article-title":"A comprehensive analysis of supermere, exomere, and extracellular vesicle isolation and cargo in colorectal cancer","volume":"44","author":"Tutanov","year":"2025","journal-title":"Cell Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"116460","DOI":"10.1016\/j.celrep.2025.116460","article-title":"Identification of molecular markers and exploration of the oncogenic role of exomeres in hepatocellular carcinoma","volume":"44","author":"Yeung","year":"2025","journal-title":"Cell Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e12470","DOI":"10.1002\/jev2.12470","article-title":"Optimized AF4 combined with density cushion ultracentrifugation enables profiling of high-purity human blood extracellular vesicles","volume":"13","author":"Hu","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"44386","DOI":"10.1021\/acsami.4c03869","article-title":"Improving the Purity of Extracellular Vesicles by Removal of Lipoproteins from Size Exclusion Chromatography- and Ultracentrifugation-Processed Samples Using Glycosaminoglycan-Functionalized Magnetic Beads","volume":"16","author":"Chou","year":"2024","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1186\/s40364-026-00947-7","article-title":"The actively secreted plasma extracellular vesicle troponin (ASPECT) study: Circulating troponin in extracellular vesicles across cardiovascular disease cohorts","volume":"14","author":"Spanos","year":"2026","journal-title":"Biomark. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3533","DOI":"10.1038\/s41467-025-57908-1","article-title":"Vesicular and non-vesicular extracellular small RNAs direct gene silencing in a plant-interacting bacterium","volume":"16","author":"Ravet","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s12929-022-00837-8","article-title":"Recent advances in conventional and unconventional vesicular secretion pathways in the tumor microenvironment","volume":"29","author":"Kuo","year":"2022","journal-title":"J. Biomed. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e12233","DOI":"10.1002\/jev2.12233","article-title":"Function of ceramide transfer protein for biogenesis and sphingolipid composition of extracellular vesicles","volume":"11","author":"Crivelli","year":"2022","journal-title":"J. Extracell. Vesicles"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2686468","DOI":"10.1080\/22221751.2026.2686468","article-title":"Extracellular vesicles participate in proteostasis and heat shock adaptation in Plasmodium falciparum","volume":"15","author":"Arce","year":"2026","journal-title":"Emerg. Microbes Infect."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"218457","DOI":"10.1016\/j.canlet.2026.218457","article-title":"Melanoma cells release dysfunctional mitochondria to the tumor microenvironment and circulation in association with tumor progression","volume":"647","author":"Fuentes","year":"2026","journal-title":"Cancer Lett."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"D1807","DOI":"10.1093\/nar\/gkaf1189","article-title":"SVAtlas: A comprehensive single extracellular vesicle omics resource","volume":"54","author":"Wei","year":"2026","journal-title":"Nucleic Acids Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"D1694","DOI":"10.1093\/nar\/gkad1007","article-title":"Vesiclepedia 2024: An extracellular vesicles and extracellular particles repository","volume":"52","author":"Chitti","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e12339","DOI":"10.1002\/jev2.12339","article-title":"Depletion of soluble cytokines unlocks the immunomodulatory bioactivity of extracellular vesicles","volume":"12","author":"Roux","year":"2023","journal-title":"J. Extracell. Vesicles"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1021\/acssensors.3c02070","article-title":"Effect of Sample Preprocessing and Size-Based Extraction Methods on the Physical and Molecular Profiles of Extracellular Vesicles","volume":"9","author":"Alexandre","year":"2024","journal-title":"ACS Sens."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"20417314231174609","DOI":"10.1177\/20417314231174609","article-title":"Comparison of extracellular vesicle isolation processes for therapeutic applications","volume":"14","author":"Williams","year":"2023","journal-title":"J. Tissue Eng."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"6681","DOI":"10.2147\/IJN.S328325","article-title":"An Isolation System to Collect High Quality and Purity Extracellular Vesicles from Serum","volume":"16","author":"Yang","year":"2021","journal-title":"Int. J. Nanomed."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e70037","DOI":"10.1002\/jev2.70037","article-title":"A Non-Centrifugation Method to Concentrate and Purify Extracellular Vesicles Using Superabsorbent Polymer Followed by Size Exclusion Chromatography","volume":"14","author":"Bergqvist","year":"2025","journal-title":"J. Extracell. Vesicles"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e70856","DOI":"10.1002\/alz70856_098434","article-title":"Comparative Analysis of Plasma-Derived Extracellular Vesicles Isolation Methods in the Context of Neurodegenerative Diseases","volume":"21","author":"Franco","year":"2025","journal-title":"Alzheimer\u2019s Dement."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3840","DOI":"10.1021\/acs.analchem.2c05352","article-title":"Development of Two-Dimensional Offline Coupling of Asymmetrical Flow Field-Flow Fractionation and Capillary Electrophoresis for the Separation of a Five-Component Submicrometer Particle Mixture","volume":"95","author":"Jing","year":"2023","journal-title":"Anal. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"e2503638","DOI":"10.1002\/smll.202503638","article-title":"Decoding Complex Biological Milieus: SHINER\u2019s Approach to Profiling and Functioning of Extracellular Vesicle Subpopulations","volume":"21","author":"Hsu","year":"2025","journal-title":"Small"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2421","DOI":"10.1039\/D2LC01193J","article-title":"Continuous-flow label-free size fractionation of extracellular vesicles through electrothermal fluid rolls and dielectrophoresis synergistically integrated in a microfluidic device","volume":"23","author":"Bu","year":"2023","journal-title":"Lab Chip"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"7212","DOI":"10.1021\/acs.analchem.4c00795","article-title":"Magnetic Nanoparticle-Based Microfluidic Platform for Automated Enrichment of High-Purity Extracellular Vesicles","volume":"96","author":"Guo","year":"2024","journal-title":"Anal. Chem."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"16759","DOI":"10.1021\/acs.analchem.5c01168","article-title":"Advanced Extracellular Vesicle Isolation: A Hybrid Electrokinetic-Tangential Flow Filtration Approach for Improved Yield, Purity, and Scalability","volume":"97","author":"Kim","year":"2025","journal-title":"Anal. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"9388","DOI":"10.1021\/acsnano.3c01340","article-title":"A Scalable High-Throughput Isoelectric Fractionation Platform for Extracellular Nanocarriers: Comprehensive and Bias-Free Isolation of Ribonucleoproteins from Plasma, Urine, and Saliva","volume":"17","author":"Sharma","year":"2023","journal-title":"ACS Nano"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Choe, S.W., Kim, B., and Kim, M. (2021). Progress of Microfluidic Continuous Separation Techniques for Micro-\/Nanoscale Bioparticles. Biosensors, 11.","DOI":"10.3390\/bios11110464"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/bs.acc.2026.03.004","article-title":"Advances in microfluidic extracellular vesicle technology","volume":"134","author":"Eigenfeld","year":"2026","journal-title":"Adv. Clin. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"e2411278","DOI":"10.1002\/advs.202411278","article-title":"Microfluidic Nanoparticle Separation for Precision Medicine","volume":"12","author":"Lan","year":"2025","journal-title":"Adv. Sci."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2861","DOI":"10.1039\/D3LC01097J","article-title":"Toward microfluidic continuous-flow and intelligent downstream processing of biopharmaceuticals","volume":"24","author":"Sharma","year":"2024","journal-title":"Lab Chip"},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Guzowska, J., Kowalski, S., Schachta, I., Pieku\u015b-S\u0142omka, N., and S\u0142omka, A. (2026). Connecting the Dots: Milestones in the History of Extracellular Vesicle Research. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27052470"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1038\/s41592-020-01034-x","article-title":"Exosome detection via the ultrafast-isolation system: EXODUS","volume":"18","author":"Chen","year":"2021","journal-title":"Nat. Methods"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.1039\/D5AN01337B","article-title":"Pinched-flow fractionation-based extracellular vesicle isolation by ExoFAST and its analytical benchmarking with commercial technologies","volume":"151","author":"Chelakkot","year":"2026","journal-title":"Analyst"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"24641","DOI":"10.3402\/jev.v3.24641","article-title":"Routes and mechanisms of extracellular vesicle uptake","volume":"3","author":"Mulcahy","year":"2014","journal-title":"J. Extracell. Vesicles"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/nrm.2017.125","article-title":"Shedding light on the cell biology of extracellular vesicles","volume":"19","author":"Raposo","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"168","DOI":"10.14336\/AD.2024.0206-1","article-title":"Nanomaterials in Medicine: Understanding Cellular Uptake, Localization, and Retention for Enhanced Disease Diagnosis and Therapy","volume":"16","author":"Peng","year":"2025","journal-title":"Aging Dis."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"35985","DOI":"10.1021\/acsami.4c04148","article-title":"Cellular Uptake of Upconversion Nanoparticles Based on Surface Polymer Coatings and Protein Corona","volume":"16","author":"Malhotra","year":"2024","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"16674","DOI":"10.1021\/acsnano.4c01640","article-title":"Enhanced Prostate-specific Membrane Antigen Targeting by Precision Control of DNA Scaffolded Nanoparticle Ligand Presentation","volume":"18","author":"Jana","year":"2024","journal-title":"ACS Nano"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"e2500903","DOI":"10.1002\/smll.202500903","article-title":"The Internal Nanostructure of Lipid Nanoparticles Influences Their Diverse Cellular Uptake Pathways","volume":"21","author":"Yap","year":"2025","journal-title":"Small"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"17069","DOI":"10.1021\/acsami.3c14592","article-title":"Extracellular Microvesicles Modified with Arginine-Rich Peptides for Active Macropinocytosis Induction and Delivery of Therapeutic Molecules","volume":"16","author":"Morimoto","year":"2024","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"115628","DOI":"10.1016\/j.colsurfb.2026.115628","article-title":"Bimodal silica nanoparticles co-doped with Mn2+ and Ru2+ complexes, variously conjugated with folates, for targeting cells and high hydrophilicity","volume":"264","author":"Bochkova","year":"2026","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"51","DOI":"10.20517\/evcna.2025.151","article-title":"Extracellular vesicles as modulators of cancer metabolism and microenvironment","volume":"7","author":"Feltri","year":"2026","journal-title":"Extracell. Vesicles Circ. Nucleic Acids"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2147\/IJN.S585732","article-title":"Bidirectional Interactions Between Nanomaterials and Extracellular Vesicles: Molecular Insights and Translational Implications","volume":"21","author":"Sa","year":"2026","journal-title":"Int. J. Nanomed."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"8046","DOI":"10.1039\/D4NR00345D","article-title":"Oral exosome-like nanovesicles from Phellinus linteus suppress metastatic hepatocellular carcinoma by reactive oxygen species generation and microbiota rebalancing","volume":"16","author":"Zu","year":"2024","journal-title":"Nanoscale"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/s12964-023-01292-0","article-title":"Extracellular vesicle therapy for obesity-induced NAFLD: A comprehensive review of current evidence","volume":"22","author":"Zhang","year":"2024","journal-title":"Cell Commun. Signal. CCS"},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Malhotra, S., Miras, M.C.M., Pappolla, A., Montalban, X., and Comabella, M. (2023). Liquid Biopsy in Neurological Diseases. Cells, 12.","DOI":"10.20944\/preprints202306.1940.v1"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"8625","DOI":"10.1021\/acs.analchem.4c05407","article-title":"Liquid Biopsy on Microfluidics: From Existing Endogenous to Emerging Exogenous Biomarkers Analysis","volume":"97","author":"Ou","year":"2025","journal-title":"Anal. Chem."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s12943-022-01509-9","article-title":"Exosomes as a new frontier of cancer liquid biopsy","volume":"21","author":"Yu","year":"2022","journal-title":"Mol. Cancer"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"122122","DOI":"10.1016\/j.ijpharm.2022.122122","article-title":"Gold nanoparticles for skin drug delivery","volume":"625","author":"Chen","year":"2022","journal-title":"Int. J. Pharm."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"798","DOI":"10.1007\/s13346-024-01681-z","article-title":"Application of quality by design in optimization of nanoformulations: Principle, perspectives and practices","volume":"15","author":"Birla","year":"2025","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1016\/j.jconrel.2021.12.024","article-title":"Emerging prospects of extracellular vesicles for brain disease theranostics","volume":"341","author":"Wang","year":"2022","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"113562","DOI":"10.1016\/j.jconrel.2025.02.058","article-title":"Nanobody-based drug delivery systems for cancer therapy","volume":"381","author":"Liu","year":"2025","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_80","first-page":"100318","article-title":"Neutrophils-mediated accelerated blood clearance phenomenon in beagles and rats based on the cross-injection of non-PEGylated and PEGylated nanoemulsions","volume":"9","author":"Liu","year":"2025","journal-title":"Int. J. Pharm. X"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"e2404860","DOI":"10.1002\/advs.202404860","article-title":"Loading of Extracellular Vesicles with Nucleic Acids via Hybridization with Non-Lamellar Liquid Crystalline Lipid Nanoparticles","volume":"12","author":"Bader","year":"2025","journal-title":"Adv. Sci."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"e2503807","DOI":"10.1002\/smll.202503807","article-title":"Droplet Squeeze Microfluidic Platform for Generating Extracellular Vesicle Hybrids for Drug Delivery","volume":"21","author":"Chintapula","year":"2025","journal-title":"Small"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"e2300319","DOI":"10.1002\/adhm.202300319","article-title":"A Comparison of Cellular Uptake Mechanisms, Delivery Efficacy, and Intracellular Fate between Liposomes and Extracellular Vesicles","volume":"12","author":"Gandek","year":"2023","journal-title":"Adv. Healthc. Mater."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"17432","DOI":"10.1021\/acs.analchem.4c04607","article-title":"Saponin is Essential for the Isolation of Proteins and RNA from Biological Nanoparticles","volume":"96","author":"Brezgin","year":"2024","journal-title":"Anal. Chem."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"2827","DOI":"10.1515\/nanoph-2022-0057","article-title":"Recent advances in optical label-free characterization of extracellular vesicles","volume":"11","author":"Imanbekova","year":"2022","journal-title":"Nanophotonics"},{"key":"ref_86","unstructured":"Ahmad, P., Farshidfar, N., Estrin, N.E., Wang, Y., Zhang, Y., Sculean, A., and Miron, R.J. (Periodontol. 2000, 2026). Understanding extracellular vesicle biology, isolation, and characterization: The theory of EV-erything, Periodontol. 2000, advance online publication."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1038\/s44319-025-00620-4","article-title":"An Argonaute protein traffics from nematode to mouse and is a vaccine against parasitic nematodes","volume":"27","author":"Neophytou","year":"2026","journal-title":"EMBO Rep."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1007\/s13346-023-01411-x","article-title":"Extracellular vesicles: Powerful candidates in nano-drug delivery systems","volume":"14","author":"Liu","year":"2024","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"2088","DOI":"10.1111\/all.16080","article-title":"Quality control of allergen products with mass spectrometry part I: Positioning within the EU regulatory framework","volume":"79","author":"Spiric","year":"2024","journal-title":"Allergy"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"e2202024","DOI":"10.1002\/smll.202202024","article-title":"Nanoparticle Tracking in Single-Antiresonant-Element Fiber for High-Precision Size Distribution Analysis of Mono- and Polydisperse Samples","volume":"18","author":"Nissen","year":"2022","journal-title":"Small"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"1654","DOI":"10.1021\/acs.analchem.4c04614","article-title":"Single Extracellular VEsicle Nanoscopy-Universal Protocol (SEVEN-UP): Accessible Imaging Platform for Quantitative Characterization of Single Extracellular Vesicles","volume":"97","author":"Saftics","year":"2025","journal-title":"Anal. Chem."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"108122","DOI":"10.1016\/j.biotechadv.2023.108122","article-title":"Technological aspects of manufacturing and analytical control of biological nanoparticles","volume":"64","author":"Brezgin","year":"2023","journal-title":"Biotechnol. Adv."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"e02394","DOI":"10.1002\/smtd.202502394","article-title":"Direct Measurement of Protein Corona Thickness on Nanomaterials Using Dual-Color Stochastic Optical Reconstruction Microscopy (STORM)","volume":"10","author":"Cech","year":"2026","journal-title":"Small Methods"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1038\/s41377-024-01623-x","article-title":"One-dimensional photonic crystal enhancing spin-to-orbital angular momentum conversion for single-particle tracking","volume":"13","author":"Huang","year":"2024","journal-title":"Light Sci. Appl."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"1831","DOI":"10.1021\/acs.nanolett.5c05825","article-title":"Label-Free Nanoparticle Characterization via Electrical Impedance-Based Submicrofluidic Analysis","volume":"26","author":"Zhao","year":"2026","journal-title":"Nano Lett."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"e2400533","DOI":"10.1002\/advs.202400533","article-title":"Addressing Heterogeneity in Direct Analysis of Extracellular Vesicles and Their Analogs by Membrane Sensing Peptides as Pan-Vesicular Affinity Probes","volume":"11","author":"Gori","year":"2024","journal-title":"Adv. Sci."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"26568","DOI":"10.1021\/acsnano.4c03679","article-title":"Simultaneous Protein and RNA Analysis in Single Extracellular Vesicles, Including Viruses","volume":"18","author":"Troyer","year":"2024","journal-title":"ACS Nano"},{"key":"ref_98","first-page":"414","article-title":"Quantum machine learning-based electrokinetic mining for the identification of nanoparticles and exosomes with minimal training data","volume":"51","author":"Thakur","year":"2025","journal-title":"Bioact. Mater."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"15375","DOI":"10.1021\/acs.analchem.3c03297","article-title":"High Confidence Single Particle Analysis with Machine Learning","volume":"95","author":"Wu","year":"2023","journal-title":"Anal. Chem."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/s12575-024-00243-4","article-title":"Comparing the Proteomic Profiles of Extracellular Vesicles Isolated using Different Methods from Long-term Stored Plasma Samples","volume":"26","author":"Torres","year":"2024","journal-title":"Biol. Proced. Online"},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"e2307240","DOI":"10.1002\/smll.202307240","article-title":"Chemically-Induced Lipoprotein Breakdown for Improved Extracellular Vesicle Purification","volume":"20","author":"Iannotta","year":"2024","journal-title":"Small"},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"13258","DOI":"10.1021\/jacs.4c17948","article-title":"High-Throughput Extracellular Vesicle Isolation Using Plate-Based Size Exclusion Chromatography and Automation","volume":"147","author":"Gilboa","year":"2025","journal-title":"J. Am. Chem. Soc."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"e04335","DOI":"10.1002\/smll.202504335","article-title":"An Integrated Digital Microfluidic Device for the Extraction and Detection of Extracellular Vesicle-Based Molecules","volume":"21","author":"Lo","year":"2025","journal-title":"Small"},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"17972","DOI":"10.1021\/acs.est.6c02876","article-title":"Attribute-Driven Maternal-Fetal Transfer of Atmospheric Magnetite Nanoparticles Revealed by Single-Particle Analysis","volume":"60","author":"Wang","year":"2026","journal-title":"Environ. Sci. Technol."},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"e12523","DOI":"10.1002\/jev2.12523","article-title":"Snorkel-tag based affinity chromatography for recombinant extracellular vesicle purification","volume":"13","author":"Bobbili","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"124921","DOI":"10.1016\/j.ijpharm.2024.124921","article-title":"Enhanced quantification and cell tracking of dual fluorescent labeled extracellular vesicles","volume":"667","author":"Leivar","year":"2024","journal-title":"Int. J. Pharm."},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"zqaf005","DOI":"10.1093\/function\/zqaf005","article-title":"Mapping Organism-wide Single Cell mRNA Expression Linked to Extracellular Vesicle Biogenesis, Secretion, and Cargo","volume":"6","author":"LaRocca","year":"2025","journal-title":"Function"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/9\/1330\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T08:49:26Z","timestamp":1789375766000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/9\/1330"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,13]]},"references-count":107,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["biom16091330"],"URL":"https:\/\/doi.org\/10.3390\/biom16091330","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,13]]}}}