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Multi-marker profiling of extracellular vesicles using streaming current and sequential electrostatic labeling
KTH, School of Engineering Sciences (SCI), Applied Physics.
Uppsala Univ, Dept Elect Engn, Div Solid State Elect, S-75121 Uppsala, Sweden..
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science.ORCID iD: 0000-0001-7755-2661
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science.ORCID iD: 0000-0002-4164-166x
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2023 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 227, article id 115142Article in journal (Refereed) Published
Abstract [en]

High heterogeneity in the membrane protein expression of small extracellular vesicles (sEVs) means that bulk methods relying on antibody-based capture for expression analysis have a drawback that each type of antibody may capture a different sub-population. An improved approach is to capture a representative sEV population, without any bias, and then perform a multiplexed protein expression analysis on this population. However, such a possibility has been largely limited to fluorescence-based methods. Here, we present a novel electrostatic labelling strategy and a microchip-based all-electric method for membrane protein analysis of sEVs. The method allows us to profile multiple surface proteins on the captured sEVs using alternating charge labels. It also permits the comparison of expression levels in different sEV-subtypes. The proof of concept was tested by capturing sEVs both non-specifically (unbiased) as well as via anti-CD9 capture probes (biased), and then profiling the expression levels of various surface proteins using the charge labelled antibodies. The method is the first of its kind, demonstrating an all-electrical and microchip based method that allows for unbiased analysis of sEV membrane protein expression, comparison of expression levels in different sEV subsets, and fractional estimation of different sEV sub-populations. These results were also validated in parallel using a single-sEV fluorescence technique.

Place, publisher, year, edition, pages
Elsevier BV , 2023. Vol. 227, article id 115142
Keywords [en]
Biosensors, Extracellular vesicles, Streaming current, Electrostatic labels, Microfluidics
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-325236DOI: 10.1016/j.bios.2023.115142ISI: 000946918000001PubMedID: 36805937Scopus ID: 2-s2.0-85148332172OAI: oai:DiVA.org:kth-325236DiVA, id: diva2:1748414
Note

QC 20230403

Available from: 2023-04-03 Created: 2023-04-03 Last updated: 2023-04-03Bibliographically approved

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