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Label-Free Surface Protein Profiling of Extracellular Vesicles by an Electrokinetic Sensor
KTH Royal Inst Technol, Sch Engn Sci, Dept Appl Phys, S-16440 Kista, Sweden.ORCID iD: 0000-0002-5077-3218
KTH Royal Inst Technol, Sch Engn Sci Chem Biotechnol & Hlth, Dept Prot Sci, AlbalNova Univ Ctr, S-10691 Stockholm, Sweden.
Karolinska Inst, Karolinska Univ Hosp, Dept Oncol Pathol, Theme Canc,Patient Area,Pelvis, Akad Straket 1, S-17164 Stockholm, Sweden.
Karolinska Inst, Clin Res Ctr, Dept Lab Med, S-17177 Stockholm, Sweden;Evox Therapeut Ltd, Oxford OX4 4HG, England.
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2019 (English)In: ACS SENSORS, ISSN 2379-3694, Vol. 4, no 5, p. 1399-1408Article in journal (Refereed) Published
Abstract [en]

Small extracellular vesicles (sEVs) generated from the endolysosomal system, often referred to as exosomes, have attracted interest as a suitable biomarker for cancer diagnostics, as they carry valuable biological information and reflect their cells of origin. Herein, we propose a simple and inexpensive electrical method for label-free detection and profiling of sEVs in the size range of exosomes. The detection method is based on the electrokinetic principle, where the change in the streaming current is monitored as the surface markers of the sEVs interact with the affinity reagents immobilized on the inner surface of a silica microcapillary. As a proof-of-concept, we detected sEVs derived from the non-small-cell lung cancer (NSCLC) cell line H1975 for a set of representative surface markers, such as epidermal growth factor receptor (EGFR), CD9, and CD63. The detection sensitivity was estimated to be similar to 175000 sEVs, which represents a sensor surface coverage of only 0.04%. We further validated the ability of the sensor to measure the expression level of a membrane protein by using sEVs displaying artificially altered expressions of EGFR and CD63, which were derived from NSCLC and human embryonic kidney (HEK) 293T cells, respectively. The analysis revealed that the changes in EGFR and CD63 expressions in sEVs can be detected with a sensitivity in the order of 10% and 3%, respectively, of their parental cell expressions. The method can be easily parallelized and combined with existing microfluidic-based EV isolation technologies, allowing for rapid detection and monitoring of sEVs for cancer diagnosis.

Place, publisher, year, edition, pages
2019. Vol. 4, no 5, p. 1399-1408
Keywords [en]
extracellular vesicles, electrokinetic effect, biosensor, label-free, protein profiling, cancer
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Engineering and Technology
Identifiers
URN: urn:nbn:se:uu:diva-387599DOI: 10.1021/acssensors.9b00418ISI: 000469410100034PubMedID: 31020844OAI: oai:DiVA.org:uu-387599DiVA, id: diva2:1331252
Funder
Swedish Research Council, 621-2014-5591Swedish Research CouncilSwedish Foundation for Strategic Research Available from: 2019-06-26 Created: 2019-06-26 Last updated: 2020-01-08Bibliographically approved

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Sahu, Siddharth S.Gatty, Hithesh KumarDev, Apurba

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