Journal of Extracellular Vesicles最新文献

{"title":"Challenges of MS-based small extracellular vesicles proteomics","authors":"Daniel Fochtman,&nbsp;Lukasz Marczak,&nbsp;Monika Pietrowska,&nbsp;Anna Wojakowska","doi":"10.1002/jev2.70020","DOIUrl":"10.1002/jev2.70020","url":null,"abstract":"<p>Proteomic profiling of small extracellular vesicles (sEV) is a powerful tool for discovering biomarkers of various diseases. This process most often assisted by mass spectrometry (MS) usually lacks standardization and recognition of challenges which may lead to unreliable results. General recommendations for sEV MS analyses have been briefly given in the MISEV2023 guidelines. The present work goes into detail for every step of sEV protein profiling with an overview of factors influencing such analyses. This includes reporting and defining the sEV source and vesicle isolation, protein solubilization and digestion, ‘offline’ and ‘online’ sample complexity reduction, the analysis type itself, and subsequent data analysis. Every stage in this process affects the others, which could result in different outcomes. Although characterization and comparisons of different sEV isolation methods are known and accessible and MS-based profiling details are provided for cell or tissue samples, no consensus work has been ever published to describe the whole process of sEV proteomic analysis. Reliable results can be obtained from sEV profiling provided that the analysis is well planned, prepared for, and backed by pilot studies or appropriate research.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142846789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"EV-Elute: A universal platform for the enrichment of functional surface marker-defined extracellular vesicle subpopulations","authors":"Willemijn S. de Voogt,&nbsp;Rowan Frunt,&nbsp;Raul M. Leandro,&nbsp;Casper S. Triesscheijn,&nbsp;Bella Monica,&nbsp;Ioanna Paspali,&nbsp;Mark Tielemans,&nbsp;Jerney J. J. M. François,&nbsp;Cor W. Seinen,&nbsp;Olivier G. de Jong,&nbsp;Sander A. A. Kooijmans","doi":"10.1002/jev2.70017","DOIUrl":"10.1002/jev2.70017","url":null,"abstract":"<p>Intercellular communication via extracellular vesicles (EVs) has been identified as a vital component of a steadily expanding number of physiological and pathological processes. To accommodate these roles, EVs have highly heterogeneous molecular compositions. Given that surface molecules on EVs determine their interactions with their environment, EV functionality likely differs between subpopulations with varying surface compositions. However, it has been technically challenging to examine such functional heterogeneity due to a lack of non-destructive methods to separate EV subpopulations based on their surface markers. Here, we used the Design-of-Experiments (DoE) methodology to optimize a protocol, which we name ‘EV-Elute’, to elute intact EVs from commercially available Protein G-coated magnetic beads. We captured EVs from various cell types on these beads using antibodies against CD9, CD63, CD81 and a custom-made protein binding phosphatidylserine (PS). When applying EV-Elute, over 70% of bound EVs could be recovered from the beads in a pH- and incubation-time-dependent fashion. EV subpopulations showed intact integrity by electron microscopy and Proteinase K protection assays and showed uptake patterns similar to whole EV isolates in co-cultures of peripheral blood mononuclear cells (PBMCs) and endothelial cells. However, in Cas9/sgRNA delivery assays, CD63⁺ EVs showed a lower capacity to functionally deliver cargo as compared to CD9⁺, CD81⁺ and PS⁺ EVs. Taken together, we developed a novel, easy-to-use platform to isolate and functionally compare surface marker-defined EV subpopulations. This platform does not require specialized equipment or reagents and is universally applicable to any capturing antibody and EV source. Hence, EV-Elute can open new opportunities to study EV functionality at the subpopulation level.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70017","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142846798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CD7-targeting pro-apoptotic extracellular vesicles: A novel approach for T-cell haematological malignancy therapy","authors":"Bei Zhang,&nbsp;Jianqiang Chen,&nbsp;Jiming Chen,&nbsp;Yingying Shen,&nbsp;Yinghu Chen,&nbsp;Shibo Wang,&nbsp;Chengyan Zhang,&nbsp;Yuzhou He,&nbsp;Huajun Feng,&nbsp;Jiaoli Wang,&nbsp;Zhijian Cai","doi":"10.1002/jev2.70025","DOIUrl":"10.1002/jev2.70025","url":null,"abstract":"<p>T-cell haematological malignancies progress rapidly and have a high mortality rate and effective treatments are still lacking. Here, we developed a drug delivery system utilizing 293T cell-derived extracellular vesicles (EVs) modified with an anti-CD7 single-chain variable fragment (αCD7/EVs). Given the challenges of chemotherapy resistance in patients with T-cell malignancy, we selected cytochrome C (CytC) and <i>Bcl2</i> siRNA (<i>siBcl2</i>) as therapeutic agents and loaded them into αCD7/EVs (αCD7/EVs/CytC/<i>siBcl2</i>). We found that αCD7/EVs efficiently targeted and were internalized by human T-ALL Molt-4 cells. In addition, the interaction between αCD7 and CD7 switched the EV entry pathway in Molt-4 cells from macropinocytosis-dependent endocytosis to clathrin-mediated endocytosis, thereby reducing EV-lysosome colocalization, ultimately improving CytC delivery efficiency and increasing the cytotoxicity of nascent EVs from EV-treated Molt-4 cells. Notably, αCD7/EVs/CytC/<i>siBcl2</i> demonstrated similar efficacy against both Molt-4 and chemotherapy-resistant Molt-4 cells (CR-Molt-4). Furthermore, αCD7/EVs/CytC/<i>siBcl2</i> exhibited high safety, low immunogenicity and minimal impact on human T cells. Therefore, αCD7/EVs/CytC/<i>siBcl2</i> are promising therapeutic approaches for treating CD7⁺ T-cell malignancies.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70025","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142828869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A decision-making tool for navigating extracellular vesicle research and product development","authors":"Francesca Loria,&nbsp;Sabrina Picciotto,&nbsp;Giorgia Adamo,&nbsp;Andrea Zendrini,&nbsp;Samuele Raccosta,&nbsp;Mauro Manno,&nbsp;Paolo Bergese,&nbsp;Giovanna L. Liguori,&nbsp;Antonella Bongiovanni,&nbsp;Nataša Zarovni","doi":"10.1002/jev2.70021","DOIUrl":"10.1002/jev2.70021","url":null,"abstract":"<p>Due to their intercellular communication properties and involvement in a wide range of biological processes, extracellular vesicles (EVs) are increasingly being studied and exploited for different applications. Nevertheless, their complex nature and heterogeneity, as well as the challenges related to their purification and characterization procedures, require a cautious assessment of the qualitative and quantitative parameters that need to be monitored. This translates into a multitude of choices and putative solutions that any EV researcher must confront in both research and translational environments. In this respect, decision-making tools may help assess various options, weigh pros and cons, and ultimately arrive at a thought-out decision that considers both the best fit-to-source and fit-to-scope EV application(s). Here, we present a multi-criteria EV decision-making grid (EV-DMG) as a novel, efficient, customizable, and easy-to-use tool to support EV research and innovation. By identifying and weighing key assessment criteria for comparing distinct EV-based preparations and related processes, our EV-DMG may assist any EV community member in making informed, traceable, and reproducible decisions regarding the management of EV sources or samples. Ultimately, this EV-DMG may guide the adoption of the most suitable EV production and analytical pipelines for targeting a defined aim or application.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11638734/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142818295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to “Beyond basic characterization and omics: Immunomodulatory roles of platelet-derived extracellular vesicles unveiled by functional testing”","authors":"","doi":"10.1002/jev2.70015","DOIUrl":"10.1002/jev2.70015","url":null,"abstract":"<p>Palviainen, M., Puutio, J., Østergaard, R. H., Eble, J. A., Maaninka, K., Butt, U., Ndika, J., Kari, O. K., Kamali-Moghaddam, M., Kjaer-Sorensen, K., Oxvig, C., Aransay, A. M., Falcon-Perez, J. M., Federico, A., Greco, D., Laitinen, S., Hayashi, Y., &amp; Siljander, P. R.-M. (2024). Beyond basic characterization and omics: Immunomodulatory roles of platelet-derived extracellular vesicles unveiled by functional testing. <i>Journal of Extracellular Vesicles</i>, <i>13</i>, e12513. https://doi.org/10.1002/jev2.12513</p><p></p><p>In Figure 1(f), the 14 hpi column showed the same image in the first row and the second row, whereas it should instead show a high contrast image of the first row following a different colour scheme as correctly shown in the other three columns (1, 4 and 7 hpi). This has been corrected by replacing the image in question. This does not affect the scientific content or the conclusion, since the results have already been fully presented in the first row of images, whereas the second row shows the same results with visual enhancement to highlight two of the three colours for clarity. There was also a typo in the label for the second row “(High constrast)”, which should have read “(High contrast)”.</p><p>In Figure 1(a), in the bright-field image, the scale bar was not fully shown. This has also been corrected by adjusting its position.</p><p>We apologize for these errors.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11638731/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142818296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Practical advice for extracellular vesicle isolation in plant–microbe interactions: Concerns, considerations, and conclusions","authors":"Hannah Thieron,&nbsp;Laura Krassini,&nbsp;Seomun Kwon,&nbsp;Sebastian Fricke,&nbsp;Sabrine Nasfi,&nbsp;Lorenz Oberkofler,&nbsp;Alessa Ruf,&nbsp;Julia Kehr,&nbsp;Karl-Heinz Kogel,&nbsp;Arne Weiberg,&nbsp;Michael Feldbrügge,&nbsp;Silke Robatzek,&nbsp;Ralph Panstruga","doi":"10.1002/jev2.70022","DOIUrl":"10.1002/jev2.70022","url":null,"abstract":"<p>In recent years, extracellular vesicles (EVs) have emerged as novel key players in plant–microbe interactions. While it is immensely useful to draw on the established “minimal information for studies of extracellular vesicles” (MISEV) guidelines and precedents in mammalian systems, working with plants and their associated microbes poses specific challenges. To navigate researchers through these obstacles, we offer detailed step-by-step suggestions for those embarking on EV research in the context of plant–microbe interactions. The advice is based on recent publications and our collective experience from the diverse plant and microbe systems studied in a dedicated research consortium. We provide considerations for experimental design, optimization, quality control, and recommendations on how to increase yield, purity, and reproducibility of EV isolation. With this perspective article, we aim not only to assist researchers in our field but also to promote discussions on plant and microbe EVs in the broader EV community.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11635479/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A perspective from the National Eye Institute Extracellular Vesicle Workshop: Gaps, needs, and opportunities for studies of extracellular vesicles in vision research","authors":"Sun Young Lee,&nbsp;Mikael Klingeborn,&nbsp;Jeff W. M. Bulte,&nbsp;Daniel T. Chiu,&nbsp;Michael Chopp,&nbsp;Christopher W. Cutler,&nbsp;Saumya Das,&nbsp;Charles E. Egwuagu,&nbsp;Christie D. Fowler,&nbsp;Sarah F. Hamm-Alvarez,&nbsp;Hakho Lee,&nbsp;Yutao Liu,&nbsp;Ben Mead,&nbsp;Tara L. Moore,&nbsp;Sriram Ravindran,&nbsp;Ashok K. Shetty,&nbsp;Johan Skog,&nbsp;Kenneth W. Witwer,&nbsp;Ali R. Djalilian,&nbsp;Alissa M. Weaver","doi":"10.1002/jev2.70023","DOIUrl":"10.1002/jev2.70023","url":null,"abstract":"<p>With an evolving understanding and new discoveries in extracellular vesicle (EV) biology and their implications in health and disease, the significant diagnostic and therapeutic potential of EVs for vision research has gained recognition. In 2021, the National Eye Institute (NEI) unveiled its Strategic Plan titled ‘Vision for the Future (2021–2025),’ which listed EV research as a priority within the domain of Regenerative Medicine, a pivotal area outlined in the Plan. In alignment with this prioritization, NEI organized a workshop inviting twenty experts from within and beyond the visual system. The workshop aimed to review current knowledge in EV research and explore gaps, needs and opportunities for EV research in the eye, including EV biology and applications of EVs in diagnosis, therapy and prognosis within the visual system. This perspective encapsulates the workshop's deliberations, highlighting the current landscape and potential implications of EV research in advancing eye health and addressing visual diseases.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11635481/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Direct imaging with multidimensional labelling and high-content analysis allows quantitative categorization and characterizations of individual small extracellular vesicles and nanoparticles (sEVPs)","authors":"Simou Sun,&nbsp;Sarah J. Cox-Vázquez,&nbsp;Nam-Joon Cho,&nbsp;Guillermo C. Bazan,&nbsp;Jay T. Groves","doi":"10.1002/jev2.12520","DOIUrl":"10.1002/jev2.12520","url":null,"abstract":"<p>Small extracellular vesicles and nanoparticles (sEVPs) are cell-secreted entities with potential as diagnostic biomarkers and therapeutic vehicles. However, significant intrinsic sEVP heterogeneity impedes analysis and understanding of their composition and functions. We employ multidimensional fluorescent labelling on sEVPs, leveraging the robustness of a newly developed membrane probe—conjugated oligoelectrolytes (COEs), and conduct total internal reflection fluorescence (TIRF) microscopy on sEVP arrays. These arrays comprise single sEVPs anchored to a soft material functionalized surface with little bias. We then develop an enhanced algorithm for colocalization analysis of the multiple labels on individual sEVPs and perform deep profiling of particle content. We categorize sEVPs derived from the same cell type into seven distinct subpopulations—some vesicular whereas others non-vesicular, and we demonstrate that sEVPs from four cell types exhibit quantitatively distinguishable subpopulation distributions. Furthermore, we gain insights into specific particle features within each subpopulation, including CD63 counts, relative particle size, relative concentration of cargoes, and correlations among different cargoes. This high-content analysis reveals common cargo sorting features in sEVP subpopulations across different cell types and suggests new statistics within the sEVP inherent heterogeneity that could differentiate sEVPs from two types of cancer cells and two types of normal cells. Collectively, our study presents a robust single-sEVP characterization platform, combining high-content imaging with comprehensive analysis. This platform is poised to advance sEVP-based theranostic assays and facilitate exploration into disease-associated sEVP biogenesis and sEVP-mediated intercellular communication.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11635478/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancing research on parasitic infections: Standardized extracellular vesicle guideline","authors":"Carmen Fernandez-Becerra,&nbsp;Patricia Xander,&nbsp;Martin Olivier,&nbsp;Ana Claudia Torrecilhas","doi":"10.1002/jev2.70009","DOIUrl":"10.1002/jev2.70009","url":null,"abstract":"&lt;p&gt;Parasitic diseases (protozoan and helminths parasites) stand as a significant global health challenge, affecting over a billion people worldwide and claiming millions of lives annually. At the heart of these diseases lie parasites, which instigate a myriad of neglected tropical and infectious diseases. Despite their significant impact on public health, these diseases present challenges in diagnosis, treatment and prevention, largely due to the complex life cycles of the parasites and the intricate nature of host–parasite interactions.&lt;/p&gt;&lt;p&gt;In recent years, the emergence of extracellular vesicles (EVs) as a topic of study has revolutionized our understanding of parasitic infections. These tiny vesicles, secreted by pathogenic protozoa and helminths parasites or infected cells, engage in crucial interactions with host cells, dictating the course of infection and disease progression. These interactions encompass a spectrum of activities crucial for the parasite's survival, including facilitating infection, modulating the host immune response, enhancing host adaptability and transferring factors that confer drug resistance. As conduits of communication between parasites and hosts, EVs hold immense potential as biomarkers for asymptomatic infections and as prognostic indicators for disease outcomes post-therapy.&lt;/p&gt;&lt;p&gt;However, despite the burgeoning interest in EVs, current methodologies for isolating, sizing and characterizing these vesicles often lack the requisite rigor, standardization and quality controls. Recognizing this gap, efforts are underway to establish comprehensive standards drawn from a growing collective understanding.&lt;/p&gt;&lt;p&gt;The journey towards understanding parasite-derived EVs has been marked by collaborative efforts and interdisciplinary dialogues. Workshop titled “Cross-Organism Communication by Extracellular Vesicles: Hosts, Microbes, Parasites (https://doi.org/10.1080/20013078.2017.1407213) organized by the International Society for Extracellular Vesicles (ISEV) workshop in 2016 - São Paulo Brazil” have served as platforms for high-level scientific discussions, exploring the nature, origin, and potential applications of EVs in infectious diseases. These gatherings underscore the need for standardized protocols and techniques for the purification and characterization of EVs, taking into account the unique characteristics of each parasite species and strain.&lt;/p&gt;&lt;p&gt;Our recently published guideline in the &lt;i&gt;Journal of Extracellular Biology&lt;/i&gt; (Fernandez-Becerra et al., &lt;span&gt;2023&lt;/span&gt;), titled “Guidelines for the Purification and Characterization of Extracellular Vesicles of Parasites,” marks a big step forward in this effort. The guideline, written by 31 scientists from around the world, offers a structured framework for isolating, characterizing and investigating EVs obtained from parasite-infected cell cultures, experimental animals and patients.&lt;/p&gt;&lt;p&gt;This editorial aims to highlight the significance of the guideline and","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11621962/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced proteomic profiling of human plasma-derived extracellular vesicles through charge-based fractionation to advance biomarker discovery potential","authors":"Xianyi Su,&nbsp;Getúlio Pereira de Oliveira Júnior,&nbsp;Anne-Lise Marie,&nbsp;Michal Gregus,&nbsp;Amanda Figueroa-Navedo,&nbsp;Ionita C. Ghiran,&nbsp;Alexander R. Ivanov","doi":"10.1002/jev2.70024","DOIUrl":"10.1002/jev2.70024","url":null,"abstract":"<p>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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11621968/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}