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Extracellular Vesic...
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
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- Mitchell, Megan, I (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Ben-Dov, Iddo Z. (author)
- Hadassah Hebrew Univ, Lab Med Transcript, Med Ctr, Jerusalem, Israel.
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- Liu, Christina (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Ye, Kenny (author)
- Albert Einstein Coll Med, Dept Epidemiol & Populat Hlth, Bronx, NY 10467 USA.
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- Chow, Kar (author)
- Hackensack Univ Med Ctr, Biorepository, Hackensack, NJ USA.
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- Kramer, Yael (author)
- Hackensack Univ Med Ctr, Biorepository, Hackensack, NJ USA.
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- Gangadharan, Anju (author)
- Hackensack Univ Med Ctr, Biorepository, Hackensack, NJ USA.
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- Park, Steven (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Fitzgerald, Sean (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Ramnauth, Andrew (author)
- Weill Cornell Med, Dept Pathol & Lab Med, New York, NY USA.
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- Perlin, David S. (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Donato, Michele (author)
- Hackensack Univ Med Ctr, Biorepository, Hackensack, NJ USA.
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- Bhoy, Emily (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Doulabi, Ehsan Manouchehri (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg
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- Poulos, Michael (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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- Kamali-Moghaddam, Masood (author)
- Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab
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- Loudig, Olivier (author)
- Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA.
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Hackensack Meridian Hlth, Ctr Discovery & Innovat, Nutley, NJ USA Hadassah Hebrew Univ, Lab Med Transcript, Med Ctr, Jerusalem, Israel. (creator_code:org_t)
- 2021-06-03
- 2021
- English.
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In: Journal of Extracellular Vesicles. - : John Wiley & Sons. - 2001-3078. ; 10:8
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Abstract
Subject headings
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- 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.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Keyword
- exosome purification
- extracellular vesicles
- micro-RNA profiling
- sequencing
- TEM
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Mitchell, Megan, ...
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Ben-Dov, Iddo Z.
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Liu, Christina
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Ye, Kenny
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Chow, Kar
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Kramer, Yael
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show more...
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Gangadharan, Anj ...
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Park, Steven
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Fitzgerald, Sean
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Ramnauth, Andrew
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Perlin, David S.
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Donato, Michele
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Bhoy, Emily
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Doulabi, Ehsan M ...
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Poulos, Michael
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Kamali-Moghaddam ...
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Loudig, Olivier
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- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Cell and Molecul ...
- Articles in the publication
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Journal of Extra ...
- By the university
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Uppsala University