{"doi":"10.7554/elife.70725","title":"Framework for rapid comparison of extracellular vesicle isolation methods","abstract":"<jats:p>Extracellular vesicles (EVs) are released by all cells into biofluids and hold great promise as reservoirs of disease biomarkers. One of the main challenges in studying EVs is a lack of methods to quantify EVs that are sensitive enough and can differentiate EVs from similarly sized lipoproteins and protein aggregates. We demonstrate the use of ultrasensitive, single-molecule array (Simoa) assays for the quantification of EVs using three widely expressed transmembrane proteins: the tetraspanins CD9, CD63, and CD81. Using Simoa to measure these three EV markers, as well as albumin to measure protein contamination, we were able to compare the relative efficiency and purity of several commonly used EV isolation methods in plasma and cerebrospinal fluid (CSF): ultracentrifugation, precipitation, and size exclusion chromatography (SEC). We further used these assays, all on one platform, to improve SEC isolation from plasma and CSF. Our results highlight the utility of quantifying EV proteins using Simoa and provide a rapid framework for comparing and improving EV isolation methods from biofluids.</jats:p>","journal":"eLife","year":2021,"id":647779,"datarank":4.289811273677857,"base_score":4.955827057601261,"endowment":4.955827057601261,"self_citation_contribution":0.7433740586401892,"citation_network_contribution":3.5464372150376677,"self_endowment_contribution":0.7433740586401892,"citer_contribution":3.5464372150376677,"corpus_percentile":null,"corpus_rank":null,"citation_count":141,"citer_count":140,"citers_with_citation_signal":122,"citers_with_endowment":122,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":76189,"name":"Maia Norman","orcid":"0000-0002-9654-4904","position":1,"is_corresponding":false},{"id":76191,"name":"Roey Lazarovits","orcid":"0000-0002-4635-9700","position":2,"is_corresponding":false},{"id":76190,"name":"Wendy Trieu","orcid":null,"position":3,"is_corresponding":false},{"id":1046277,"name":"Ju-Hyun Lee","orcid":"0000-0001-6728-2071","position":4,"is_corresponding":false},{"id":157735,"name":"George M Church","orcid":null,"position":5,"is_corresponding":false},{"id":76195,"name":"David R. 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Using Simoa to measure these three EV markers, as well as albumin to measure protein contamination, we were able to compare the relative efficiency and purity of several commonly used EV isolation methods in plasma and cerebrospinal fluid (CSF): ultracentrifugation, precipitation, and size exclusion chromatography (SEC). We further used these assays, all on one platform, to improve SEC isolation from plasma and CSF. Our results highlight the utility of quantifying EV proteins using Simoa and provide a rapid framework for comparing and improving EV isolation methods from biofluids.</jats:p>","is_dataset_classified":null,"base_score":4.955827057601261,"endowment":4.955827057601261,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"34783650","pmcid":"PMC8651285","openalex_id":"https://openalex.org/W3213442857","authors":[],"funders":[{"funder_name":"Chan Zuckerberg Initiative","grant_id":"NDCN Collaborative Science Award","title":null},{"funder_name":"Fundação para a Ciência e a Tecnologia, I.P.","grant_id":"Incentivo/SAU/LA0002/2013","title":"Incentive - LA 2 - 2013"},{"funder_name":"Open Philanthropy Project","grant_id":"","title":null}],"total_grants":3,"fwci":6.2984,"citation_percentile":0.9790273,"influential_citations":0,"citation_trend":[{"year":2022,"count":17},{"year":2023,"count":32},{"year":2024,"count":36},{"year":2025,"count":38},{"year":2026,"count":18}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://doi.org/10.7554/elife.70725","host_type":"journal"},{"url":"https://doi.org/10.7554/elife.70725","host_type":"publisher"},{"url":"https://cdn.elifesciences.org/articles/70725/elife-70725-v2.pdf","host_type":"publisher"},{"url":"https://cdn.elifesciences.org/articles/70725/elife-70725-v2.xml","host_type":"publisher"},{"url":"https://elifesciences.org/articles/70725","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/34783650","host_type":"repository"},{"url":"https://doaj.org/article/890c2867abed4743ac6ffd19afe0b138","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/8651285","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC8651285","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC8651285?pdf=render","host_type":"Europe_PMC"},{"url":"https://doi.org/10.1101/2020.10.13.337881","host_type":""},{"url":"https://www.biorxiv.org/content/biorxiv/early/2021/05/26/2020.10.13.337881.full.pdf","host_type":""},{"url":"http://dx.doi.org/10.7554/eLife.70725","host_type":""},{"url":"https://dx.doi.org/10.1101/2020.10.13.337881","host_type":""},{"url":"https://dx.doi.org/10.7554/elife.70725","host_type":""}],"fields_of_study":["Extracellular vesicles in disease","Lipid Membrane Structure and Behavior","Caveolin-1 and cellular processes","0301 basic medicine","0303 health sciences","03 medical and health sciences"],"mesh_terms":["Extracellular Vesicles","Albumins","Cerebrospinal Fluid","Chromatography, Gel","Humans","Plasma","Ultracentrifugation","Tetraspanins"],"keywords":["Extracellular vesicles","Extracellular vesicle","Isolation (microbiology)","Vesicle","Albumin","Transmembrane protein","Extracellular","Analyte","Human","Biochemistry","Medicine","Biomarkers","Size exclusion chromatography","Diagnostics","Exosomes","Chemical Biology","QH301-705.5","Tetraspanins","Science","Q","R","Plasma","Biochemistry and Chemical Biology","Albumins","Chromatography, Gel","Humans","Biology (General)","Ultracentrifugation","Cerebrospinal Fluid"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"doi"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-10T01:52:07.690738Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}