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The high level of variability and low reproducibility rate of EV\/extracellular RNA (exRNA) results generated even on the same biofluids promoted several efforts to limit pre\u2010analytical variability by standardizing sample collection and sample preparation, along with instrument validation, setup and calibration. Anticoagulants (ACs) are often chosen based on the initial goal of the study and not necessarily for the later EV and\/or exRNA analyses. We show the effects of blood collection on EV size, abundance, and antigenic composition, as well on the miRNAs. Our focus of this work was on the effect of ACs on the number and antigenic composition of circulating EVs and on a set of circulating miRNA species, which were shown to be relevant as disease markers in several cancers and Alzheimer's disease. Results show that while the number of plasma EVs, their relative size, and post\u2010fluorescence labeling profile varied with each AC, their overall antigenic composition, with few exceptions, did not change significantly. However, the number of EVs expressing platelet and platelet\u2010activation markers increased in serum samples. For overall miRNA expression levels, EDTA was a better AC, although this may have been associated with stimulation of cells in the blood collection tube. Citrate and serum rendered better results for a set of miRNAs that were described as circulating markers for Alzheimer's disease, colon, and papillary thyroid cancers.<\/jats:p>","DOI":"10.1002\/jex2.70008","type":"journal-article","created":{"date-parts":[[2024,10,22]],"date-time":"2024-10-22T08:39:22Z","timestamp":1729586362000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Choice of blood collection methods influences extracellular vesicles counts and miRNA profiling"],"prefix":"10.1002","volume":"3","author":[{"given":"Vivian","family":"Tran","sequence":"first","affiliation":[{"name":"Department of Anesthesia, Beth Israel Deaconess Medical Center Harvard Medical School  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6940-401X","authenticated-orcid":false,"given":"Getulio Pereira de","family":"Oliveira\u2010Jr","sequence":"additional","affiliation":[{"name":"Division of Allergy and Inflammation, Department of Medicine, Beth Israel Deaconess Medical Center Harvard Medical School  Boston Massachusetts USA"},{"name":"Department of Chemistry and Chemical Biology, The Barnett Institute of Chemical &amp; Biological Analysis Northeastern University  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanie","family":"Chidester","sequence":"additional","affiliation":[{"name":"Laboratory of Pathology, Center for Cancer Research National Cancer Institute  Bethesda Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shulin","family":"Lu","sequence":"additional","affiliation":[{"name":"Division of Allergy and Inflammation, Department of Medicine, Beth Israel Deaconess Medical Center Harvard Medical School  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michelle L.","family":"Pleet","sequence":"additional","affiliation":[{"name":"Neuroimmunology and Neurovirology Division, National Institute for Neurological Disease and Stroke National Institutes of Health  Bethesda Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander R.","family":"Ivanov","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Chemical Biology, The Barnett Institute of Chemical &amp; Biological Analysis Northeastern University  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Tigges","sequence":"additional","affiliation":[{"name":"Nanoflow Cytometry Core Facility, Beth Israel Deaconess Medical Center Harvard Medical School  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moua","family":"Yang","sequence":"additional","affiliation":[{"name":"Division of Hemostasis and Thrombosis, Department of Medicine, Beth Israel Deaconess Medical Center Harvard Medical School  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven","family":"Jacobson","sequence":"additional","affiliation":[{"name":"Neuroimmunology and Neurovirology Division, National Institute for Neurological Disease and Stroke National Institutes of Health  Bethesda Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria C. 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