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Characterizing sing...
Characterizing single extracellular vesicles by droplet barcode sequencing for protein analysis
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- Banijamali, Mahsan (author)
- KTH,Genteknologi,Science for Life Laboratory, SciLifeLab
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- Höjer, Pontus (author)
- KTH,Genteknologi,Science for Life Laboratory, SciLifeLab
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- Nagy, Abel (author)
- KTH,Proteinvetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Haag, Petra (author)
- Karolinska Institutet
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- Paz Gomero, Elizabeth (author)
- KTH,Proteinvetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Stiller, Christiane (author)
- KTH,Proteinvetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Kaminskyy, Vitaliy O. (author)
- Karolinska Institutet
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- Ekman, Simon (author)
- Karolinska Institutet
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- Lewensohn, Rolf (author)
- Karolinska Inst, Dept Oncol Pathol, Solna, Sweden.;Karolinska Univ Hosp, Theme Canc, Med Unit Head & Neck Lung & Skin Tumors, Thorac Oncol Ctr, Solna, Sweden.
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- Eriksson Karlström, Amelie (author)
- KTH,Albanova VinnExcellence Center for Protein Technology, ProNova,Proteinvetenskap
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- Viktorsson, Kristina (author)
- Karolinska Institutet
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- Ahmadian, Afshin (author)
- KTH,Genteknologi,Science for Life Laboratory, SciLifeLab
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(creator_code:org_t)
- 2022-11-03
- 2022
- English.
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In: Journal of Extracellular Vesicles. - : Wiley. - 2001-3078. ; 11:11
- Related links:
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Subject headings
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- Small extracellular vesicles (sEVs) have in recent years evolved as a source of biomarkers for disease diagnosis and therapeutic follow up. sEV samples derived from multicellular organisms exhibit a high heterogeneous repertoire of vesicles which current methods based on ensemble measurements cannot capture. In this work we present droplet barcode sequencing for protein analysis (DBS-Pro) to profile surface proteins on individual sEVs, facilitating identification of sEV-subtypes within and between samples. The method allows for analysis of multiple proteins through use of DNA barcoded affinity reagents and sequencing as readout. High throughput single vesicle profiling is enabled through compartmentalization of individual sEVs in emulsion droplets followed by droplet barcoding through PCR. In this proof-of-concept study we demonstrate that DBS-Pro allows for analysis of single sEVs, with a mixing rate below 2%. A total of over 120,000 individual sEVs obtained from a NSCLC cell line and from malignant pleural effusion (MPE) fluid of NSCLC patients have been analyzed based on their surface proteins. We also show that the method enables single vesicle surface protein profiling and by extension characterization of sEV-subtypes, which is essential to identify the cellular origin of vesicles in heterogenous samples.
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
- droplet barcode sequencing (DBS)
- droplet barcode sequencing for protein analysis (DBS-Pro)
- protein profiling
- sEV subtypes
- single vesicle
- small extracellular vesicles (sEVs)
- surface protein
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Banijamali, Mahs ...
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Höjer, Pontus
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Nagy, Abel
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Haag, Petra
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Paz Gomero, Eliz ...
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Stiller, Christi ...
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Kaminskyy, Vital ...
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Ekman, Simon
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Lewensohn, Rolf
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Eriksson Karlstr ...
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Viktorsson, Kris ...
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Ahmadian, Afshin
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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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Royal Institute of Technology
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Karolinska Institutet