Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential
Abstract The study introduces a charge‐based fractionation method for fractionating plasma‐derived extracellular vesicles (EVs) into sub‐populations aimed at the improved purification from free plasma proteins to enhance the diagnostic potential of EV sub‐populations for specific pathophysiological...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2024-12-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jev2.70024 |
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| author | Xianyi Su Getúlio Pereira de Oliveira Júnior Anne‐Lise Marie Michal Gregus Amanda Figueroa‐Navedo Ionita C. Ghiran Alexander R. Ivanov |
| author_facet | Xianyi Su Getúlio Pereira de Oliveira Júnior Anne‐Lise Marie Michal Gregus Amanda Figueroa‐Navedo Ionita C. Ghiran Alexander R. Ivanov |
| author_sort | Xianyi Su |
| collection | DOAJ |
| description | Abstract The study introduces a charge‐based fractionation method for fractionating plasma‐derived extracellular vesicles (EVs) into sub‐populations aimed at the improved purification from free plasma proteins to enhance the diagnostic potential of EV sub‐populations for specific pathophysiological states. Here, we present a novel approach for EV fractionation that leverages EVs’ inherent surface charges, differentiating them from other plasma components and, thus, reducing the sample complexity and increasing the purity of EVs. The developed method was optimized and thoroughly evaluated using proteomic analysis, transmission electron microscopy, nanoparticle tracking, and western blotting of isolated EVs from healthy donors. Subsequently, we pilot‐tested the developed technique for its applicability to real‐world specimens using a small set of clinical prostate cancer samples and matched controls. The presented technique demonstrates the effective isolation and fractionation of EV sub‐populations based on their surface charge, which may potentially help enhance EV‐based diagnostics, biomarker discovery, and basic biology research. The method is designed to be straightforward, scalable, easy‐to‐use, and it does not require specialized skills or equipment. |
| format | Article |
| id | doaj-art-c7a78bcf58e44a778a05b1961805e408 |
| institution | Kabale University |
| issn | 2001-3078 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj-art-c7a78bcf58e44a778a05b1961805e4082025-01-17T11:11:12ZengWileyJournal of Extracellular Vesicles2001-30782024-12-011312n/an/a10.1002/jev2.70024Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potentialXianyi Su0Getúlio Pereira de Oliveira Júnior1Anne‐Lise Marie2Michal Gregus3Amanda Figueroa‐Navedo4Ionita C. Ghiran5Alexander R. Ivanov6Department of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonMassachusettsUSADepartment of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonMassachusettsUSADepartment of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonMassachusettsUSADepartment of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonMassachusettsUSADepartment of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonMassachusettsUSADepartment of Anesthesia, Beth Israel Deaconess Medical CenterHarvard Medical SchoolBostonMassachusettsUSADepartment of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonMassachusettsUSAAbstract The study introduces a charge‐based fractionation method for fractionating plasma‐derived extracellular vesicles (EVs) into sub‐populations aimed at the improved purification from free plasma proteins to enhance the diagnostic potential of EV sub‐populations for specific pathophysiological states. Here, we present a novel approach for EV fractionation that leverages EVs’ inherent surface charges, differentiating them from other plasma components and, thus, reducing the sample complexity and increasing the purity of EVs. The developed method was optimized and thoroughly evaluated using proteomic analysis, transmission electron microscopy, nanoparticle tracking, and western blotting of isolated EVs from healthy donors. Subsequently, we pilot‐tested the developed technique for its applicability to real‐world specimens using a small set of clinical prostate cancer samples and matched controls. The presented technique demonstrates the effective isolation and fractionation of EV sub‐populations based on their surface charge, which may potentially help enhance EV‐based diagnostics, biomarker discovery, and basic biology research. The method is designed to be straightforward, scalable, easy‐to‐use, and it does not require specialized skills or equipment.https://doi.org/10.1002/jev2.70024EV sub‐populationsextracellular vesiclesion exchange chromatographyprostate cancer diagnosticsproteomics |
| spellingShingle | Xianyi Su Getúlio Pereira de Oliveira Júnior Anne‐Lise Marie Michal Gregus Amanda Figueroa‐Navedo Ionita C. Ghiran Alexander R. Ivanov Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential Journal of Extracellular Vesicles EV sub‐populations extracellular vesicles ion exchange chromatography prostate cancer diagnostics proteomics |
| title | Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential |
| title_full | Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential |
| title_fullStr | Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential |
| title_full_unstemmed | Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential |
| title_short | Enhanced proteomic profiling of human plasma‐derived extracellular vesicles through charge‐based fractionation to advance biomarker discovery potential |
| title_sort | enhanced proteomic profiling of human plasma derived extracellular vesicles through charge based fractionation to advance biomarker discovery potential |
| topic | EV sub‐populations extracellular vesicles ion exchange chromatography prostate cancer diagnostics proteomics |
| url | https://doi.org/10.1002/jev2.70024 |
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