On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis
Extracellular vesicles (EVs) and their cargo are increasingly suggested as innovative biomarkers correlated to the diagnosis, progression and therapy of diseases like cancer. Several techniques have been developed for the specific separation of the different classes of EVs that give solutions enrich...
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| Format: | Article |
| Language: | English |
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MDPI AG
2025-03-01
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| Series: | Chemosensors |
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| Online Access: | https://www.mdpi.com/2227-9040/13/3/83 |
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| author | Cristina Potrich Anna Pedrotti Lia Vanzetti Cecilia Pederzolli Lorenzo Lunelli |
| author_facet | Cristina Potrich Anna Pedrotti Lia Vanzetti Cecilia Pederzolli Lorenzo Lunelli |
| author_sort | Cristina Potrich |
| collection | DOAJ |
| description | Extracellular vesicles (EVs) and their cargo are increasingly suggested as innovative biomarkers correlated to the diagnosis, progression and therapy of diseases like cancer. Several techniques have been developed for the specific separation of the different classes of EVs that give solutions enriched in vesicles, but still containing other unwanted components. New methods for a more efficient, reliable and automated isolation of EVs are therefore highly desirable. Here, microparticles with surfaces endowed with positive ions were exploited to separate vesicles from complex biological matrices. First, flat silicon oxide surfaces functionalized with different divalent cations were tested for their efficiency in terms of small EV capture. Small EVs pre-purified via serial ultracentrifugations were employed for these analyses. The two better-performing cations, i.e., Cu<sup>2+</sup> and Ni<sup>2+</sup>, were then selected to functionalize magnetic microbeads to be inserted in microfluidic chips and evaluated for their efficiency in capturing EVs and for their release of biomarkers. The best protocol setup was explored for the capture of EVs from cell culture supernatants and for the analysis of a class of biomarkers, i.e., microRNAs, via RT-PCR. The promising results obtained with this on-chip protocol evidenced the potential automation, miaturization, ease-of-use and the effective speed of the method, allowing a step forward toward its integration in simple and fast biosensors capable of analyzing the desired biomarkers present in EVs, helping the spread of biomarker analysis in both clinical settings and in research. |
| format | Article |
| id | doaj-art-36797ce4190e4640963ce42fec2da09f |
| institution | Kabale University |
| issn | 2227-9040 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Chemosensors |
| spelling | doaj-art-36797ce4190e4640963ce42fec2da09f2025-08-20T03:43:33ZengMDPI AGChemosensors2227-90402025-03-011338310.3390/chemosensors13030083On-Chip Purification of Extracellular Vesicles for microRNA Biomarker AnalysisCristina Potrich0Anna Pedrotti1Lia Vanzetti2Cecilia Pederzolli3Lorenzo Lunelli4Fondazione Bruno Kessler, Center for Sensors & Devices, Via Sommarive 18, Povo, 38123 Trento, ItalyFondazione Bruno Kessler, Center for Sensors & Devices, Via Sommarive 18, Povo, 38123 Trento, ItalyFondazione Bruno Kessler, Center for Sensors & Devices, Via Sommarive 18, Povo, 38123 Trento, ItalyFondazione Bruno Kessler, Center for Sensors & Devices, Via Sommarive 18, Povo, 38123 Trento, ItalyFondazione Bruno Kessler, Center for Sensors & Devices, Via Sommarive 18, Povo, 38123 Trento, ItalyExtracellular vesicles (EVs) and their cargo are increasingly suggested as innovative biomarkers correlated to the diagnosis, progression and therapy of diseases like cancer. Several techniques have been developed for the specific separation of the different classes of EVs that give solutions enriched in vesicles, but still containing other unwanted components. New methods for a more efficient, reliable and automated isolation of EVs are therefore highly desirable. Here, microparticles with surfaces endowed with positive ions were exploited to separate vesicles from complex biological matrices. First, flat silicon oxide surfaces functionalized with different divalent cations were tested for their efficiency in terms of small EV capture. Small EVs pre-purified via serial ultracentrifugations were employed for these analyses. The two better-performing cations, i.e., Cu<sup>2+</sup> and Ni<sup>2+</sup>, were then selected to functionalize magnetic microbeads to be inserted in microfluidic chips and evaluated for their efficiency in capturing EVs and for their release of biomarkers. The best protocol setup was explored for the capture of EVs from cell culture supernatants and for the analysis of a class of biomarkers, i.e., microRNAs, via RT-PCR. The promising results obtained with this on-chip protocol evidenced the potential automation, miaturization, ease-of-use and the effective speed of the method, allowing a step forward toward its integration in simple and fast biosensors capable of analyzing the desired biomarkers present in EVs, helping the spread of biomarker analysis in both clinical settings and in research.https://www.mdpi.com/2227-9040/13/3/83extracellular vesiclesmicrofluidic purificationbiomarkersmicroRNAsmicrobeads |
| spellingShingle | Cristina Potrich Anna Pedrotti Lia Vanzetti Cecilia Pederzolli Lorenzo Lunelli On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis Chemosensors extracellular vesicles microfluidic purification biomarkers microRNAs microbeads |
| title | On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis |
| title_full | On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis |
| title_fullStr | On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis |
| title_full_unstemmed | On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis |
| title_short | On-Chip Purification of Extracellular Vesicles for microRNA Biomarker Analysis |
| title_sort | on chip purification of extracellular vesicles for microrna biomarker analysis |
| topic | extracellular vesicles microfluidic purification biomarkers microRNAs microbeads |
| url | https://www.mdpi.com/2227-9040/13/3/83 |
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