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Series GSE158948 Query DataSets for GSE158948
Status Public on May 13, 2021
Title A customizable purification assay designed for small-RNA biomarker identification and functional evaluation of circulating exosomes: Exosome Capture by AnTibody of CHoice and Enzymatic Release (exo-CATCHER)
Organisms Homo sapiens; Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Circulating nucleic acids, encapsulated within exosomes, can provide a remote cellular snapshot of biomarkers associated with disease, however selective exosome isolation is critical. Current laboratory-based purification techniques rely on the physical properties of exosomes for selection, rather than their inherited cellular characteristics. Therefore, we established a highly selective purification assay, termed exosome Capture by AnTibody of CHoice and Enzymatic Release (exo-CATCHER), designed for high-throughput analysis of low-abundance small-RNA cargos by next-generation sequencing. We demonstrated its sensitivity by specifically isolating and sequencing small-RNA from mouse exosomes spiked into human plasma. Furthermore, we used Western blotting, nanoparticle tracking, and transmission electron microscopy to validate, quantify, and demonstrate capture and release of intact exosomes. As proof of principle, we utilized exo-CATCHER to purify serum exosomes from mildly and severely ill Covid-19 patients, identified, and validated hsa-miR-146a and hsa-miR-126-3p to be significantly downregulated in a small sample set of severely ill patients. We used exo-CATCHER to purify intact exosomes from high IgG titer convalescent sera and confirmed unexpected neutralizing properties, against SARS-CoV-2 in vitro, identified using ultracentrifuged exosomes, not observable with exosomes from sera with IgG titers below quantification level. Exo-CATCHER represents a versatile molecular assay for highly specific purification of circulating exosomes.
 
Overall design miRNA profiling of serum, plasma and antibody-captured circulating exosomes
 
Contributor(s) Mitchell M, Ben-Dov IZ, Kamali‐Moghaddam M, Loudig O
Citation(s) 34122779
Submission date Oct 02, 2020
Last update date Jun 15, 2021
Contact name Iddo Z. Ben-Dov
E-mail(s) iddo@hadassah.org.il
Phone +97226776881
Organization name Hadassah Medical Center
Department Nephrology and Hypertension
Lab Laboratory of Medical Transcriptomics
Street address Ein Kerem
City Jerusalem
ZIP/Postal code 91120
Country Israel
 
Platforms (3)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
GPL17021 Illumina HiSeq 2500 (Mus musculus)
GPL22245 Illumina HiSeq 2500 (Homo sapiens; Mus musculus)
Samples (74)
GSM4816143 Covid-19-Exo_3-26_75
GSM4816144 Covid-19-Exo_3-26_76
GSM4816145 Covid-19-Exo_3-26_77
Relations
BioProject PRJNA667083
SRA SRP286199

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE158948_lib1-lib4_merged_mir_summary.xlsx 359.5 Kb (ftp)(http) XLSX
GSE158948_merged_mir_Exo3.xlsx 117.5 Kb (ftp)(http) XLSX
GSE158948_merged_mir_Exo5B.xlsx 44.6 Kb (ftp)(http) XLSX
GSE158948_qPCR_melted_results.xlsx 48.5 Kb (ftp)(http) XLSX
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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