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Sensitive and Specific Detection of Platelet-Derived and Tissue Factor-Positive Extracellular Vesicles in Plasma using Solid-Phase Proximity Ligation Assay
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Chemistry. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Chemistry.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Cardiology.
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2018 (English)In: Thrombosis and Haemostasis open, ISSN 2512-9465Article in journal (Refereed) Published
Abstract [en]

Extracellular vesicles (EVs) derived from blood cells are promising biomarkers for various diseases. However, they are difficult to measure accurately in plasma due to their small size. Here, we demonstrate that platelet-derived EVs in plasma can be measured using solid-phase proximity ligation assay with high sensitivity and specificity using very small sample volume of biological materials. The results correlate well with high-sensitivity flow cytometry with the difference that the smallest EVs are detected. Briefly, the EVs are first captured on a solid phase, using lactadherin binding, and detection requires recognition with two antibodies followed by qPCR. The assay, using cholera toxin subunit-B or lactadherin as capture agents, also allowed detection of the more rare population of tissue factor (TF)-positive EVs at a concentration similar to sensitive TF activity assays. Thus, this assay can detect different types of EVs with high specificity and sensitivity, and has the potential to be an attractive alternative to flow cytometric analysis of preclinical and clinical samples. Improved techniques for measuring EVs in plasma will hopefully contribute to the understanding of their role in several diseases.

Place, publisher, year, edition, pages
Stuttgart: Georg Thieme Verlag KG, 2018.
Keywords [en]
microvesicles, myocardial infarction, cardiovascular diseases, vascular homeostasis
National Category
Cardiac and Cardiovascular Systems
Identifiers
URN: urn:nbn:se:uu:diva-367344DOI: 10.1055/s-0038-1667204OAI: oai:DiVA.org:uu-367344DiVA, id: diva2:1267088
Available from: 2018-11-30 Created: 2018-11-30 Last updated: 2019-02-14Bibliographically approved

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Thulin, ÅsaYan, JunhongÅberg, MikaelChristersson, ChristinaKamali-Moghaddam, MasoodSiegbahn, Agneta
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Clinical ChemistryScience for Life Laboratory, SciLifeLabDepartment of Immunology, Genetics and PathologyCardiology
Cardiac and Cardiovascular Systems

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