Addressing challenges in the removal of unbound dye from passively labelled extracellular vesicles.
| Autor: | Rautaniemi K; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Zini J; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki Viikinkaari 5 00790 Helsinki Finland., Löfman E; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Saari H; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki Viikinkaari 5 00790 Helsinki Finland.; Finnish Red Cross Blood Services Kivihaantie 7 00310 Helsinki Finland., Haapalehto I; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Laukka J; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Vesamäki S; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Efimov A; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Yliperttula M; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki Viikinkaari 5 00790 Helsinki Finland., Laaksonen T; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi.; Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki Viikinkaari 5 00790 Helsinki Finland., Vuorimaa-Laukkanen E; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi., Lisitsyna ES; Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University Korkeakoulunkatu 8 33720 Tampere Finland ekaterina.lisitsyna@tuni.fi. |
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| Jazyk: | angličtina |
| Zdroj: | Nanoscale advances [Nanoscale Adv] 2021 Nov 23; Vol. 4 (1), pp. 226-240. Date of Electronic Publication: 2021 Nov 23 (Print Publication: 2021). |
| DOI: | 10.1039/d1na00755f |
| Abstrakt: | Studies of extracellular vesicles (EVs), their trafficking and characterization often employ fluorescent labelling. Unfortunately, little attention has been paid thus far to a thorough evaluation of the purification of EVs after labelling, although the presence of an unbound dye may severely compromise the results or even lead to wrong conclusions on EV functionality. Here, we systematically studied five dyes for passive EV labelling and meticulously compared five typical purification methods: ultracentrifugation (UC), ultracentrifugation with discontinuous density gradient (UCG), ultrafiltration (UF), size exclusion chromatography (SEC), and anion exchange chromatography (AEC). A general methodology for evaluation of EV purification efficiency after the labelling was developed and tested to select the purification methods for the chosen dyes. Firstly, we found that some methods initially lead to high EV losses even in the absence of the dye. Secondly, the suitable purification method needs to be found for each particular dye and depends on the physical and chemical properties of the dye. Thirdly, we demonstrated that the developed parameter E Competing Interests: There are no conflicts to declare. (This journal is © The Royal Society of Chemistry.) |
| Databáze: | MEDLINE |
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