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10.1002/jev2.12110

http://scihub22266oqcxt.onion/10.1002/jev2.12110
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34122779!8173589!34122779
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suck abstract from ncbi


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pmid34122779      J+Extracell+Vesicles 2021 ; 10 (8): e12110
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  • Extracellular Vesicle Capture by AnTibody of CHoice and Enzymatic Release (EV-CATCHER): A customizable purification assay designed for small-RNA biomarker identification and evaluation of circulating small-EVs #MMPMID34122779
  • Mitchell MI; Ben-Dov IZ; Liu C; Ye K; Chow K; Kramer Y; Gangadharan A; Park S; Fitzgerald S; Ramnauth A; Perlin DS; Donato M; Bhoy E; Manouchehri Doulabi E; Poulos M; Kamali-Moghaddam M; Loudig O
  • J Extracell Vesicles 2021[Jun]; 10 (8): e12110 PMID34122779show ga
  • Circulating nucleic acids, encapsulated within small extracellular vesicles (EVs), provide a remote cellular snapshot of biomarkers derived from diseased tissues, however selective isolation is critical. Current laboratory-based purification techniques rely on the physical properties of small-EVs rather than their inherited cellular fingerprints. We established a highly-selective purification assay, termed EV-CATCHER, initially designed for high-throughput analysis of low-abundance small-RNA cargos by next-generation sequencing. We demonstrated its selectivity by specifically isolating and sequencing small-RNAs from mouse small-EVs spiked into human plasma. Western blotting, nanoparticle tracking, and transmission electron microscopy were used to validate and quantify the capture and release of intact small-EVs. As proof-of-principle for sensitive detection of circulating miRNAs, we compared small-RNA sequencing data from a subset of small-EVs serum-purified with EV-CATCHER to data from whole serum, using samples from a small cohort of recently hospitalized Covid-19 patients. We identified and validated, only in small-EVs, hsa-miR-146a and hsa-miR-126-3p to be significantly downregulated with disease severity. Separately, using convalescent sera from recovered Covid-19 patients with high anti-spike IgG titers, we confirmed the neutralizing properties, against SARS-CoV-2 in vitro, of a subset of small-EVs serum-purified by EV-CATCHER, as initially observed with ultracentrifuged small-EVs. Altogether our data highlight the sensitivity and versatility of EV-CATCHER.
  • |Animals[MESH]
  • |Bodily Secretions/chemistry[MESH]
  • |COVID-19/blood/physiopathology[MESH]
  • |Chlorocebus aethiops[MESH]
  • |Circulating MicroRNA[MESH]
  • |Extracellular Vesicles/*chemistry[MESH]
  • |High-Throughput Nucleotide Sequencing[MESH]
  • |Humans[MESH]
  • |Immunologic Techniques/*methods[MESH]
  • |MCF-7 Cells[MESH]
  • |Mice[MESH]
  • |RAW 264.7 Cells[MESH]
  • |Severity of Illness Index[MESH]


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  • suck abstract from ncbi

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