An integrated magneto-electrochemical device for the rapid profiling of tumour extracellular vesicles from blood plasma

Park, Jong-Min; Park, Jun Seok; Huang, Chen; Jo, Ala; Cook, Kaitlyn; Wang, Rui; Lin, Hsing Ying; Deun, Jan Van; Li, Yong; Min, Jouha; Wang, Lan; Yoon, Ghil-Suk; Carter, Bob S.; Balaj, Leonora; Choi, Gyu Seog; Castro, Cesar M.; Weissleder, Ralph; Lee, Hakho

doi:10.1038/s41551-021-00752-7

Cited by 111 publications

(95 citation statements)

References 62 publications

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“…We next performed bulk assays for protein and mRNA targets. For protein measurements, we used the established integrated magneto-electrochemical exosome (iMEX) platform (see Methods for details) 28 - 30 . We biotinylated lipid vesicles with amine-reactive biotin and captured them on streptavidin-coated magnetic beads.…”

Section: Resultsmentioning

confidence: 99%

Characterization and modulation of surface charges to enhance extracellular vesicle isolation in plasma

Woo¹,

Cho²,

Lee³

et al. 2022

Theranostics

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Extracellular vesicles (EVs) carry information inherited from parental cells, having significant potential for disease diagnosis. In blood, however, EVs are outnumbered >10 4 -fold by low density lipoproteins (LDLs), yet similar in size and density. These fundamental disadvantages often cause LDL spillover into EV isolates, thus confounding assay results. We hypothesized that EVs can be further separated from LDLs based on electric charge: EVs and LDLs have different lipid composition, which can lead to differential surface charge densities. To test this hypothesis, we modeled and quantified the surface charge of EVs and LDLs, and used the information to optimally separate EVs from LDLs via ion-exchange chromatography. Methods: We built an enhanced dual-mode chromatography (eDMC) device which performed i) size-exclusion to remove particles smaller than EVs and LDLs and ii) cation-exchange in an acidic elution to retain LDLs longer than EVs. The performance of the eDMC, in comparison to size-exclusion only, was evaluated by analyzing the yield and purity of the isolated EVs. Results: By measuring and modeling zeta potentials at different buffer pH, we estimated surface charge densities of EVs (-6.2 mC/m 2 ) and LDLs (-3.6 mC/m 2 ), revealing that EVs are more negatively charged than LDLs. Furthermore, the charge difference between EVs and LDLs was maximal at a weak acidic condition (pH = 6.4). By applying these findings, we optimized eDMC operation to enrich EVs directly from plasma, depleting >99.8% of LPPs within 30 min. Minimizing LDL contamination improved analytical signals in EV molecular assays, including single vesicle imaging, bulk protein measurements, and mRNA detection. Conclusions: These developments will promote the translational value of the dual-mode separation - a fast, equipment-free, and non-biased way for EV isolation from plasma samples.

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Section: Resultsmentioning

confidence: 99%

Characterization and modulation of surface charges to enhance extracellular vesicle isolation in plasma

Woo¹,

Cho²,

Lee³

et al. 2022

Theranostics

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“… Thakur et al (2021) presented a localized surface plasmon resonance (LSPR) technique to detect tumor-derived EVs using designed TiN-NH-LSPR biosensor and demonstrated that the label-free LSPR technique can be used for glioblastoma monitoring. Park et al (2021) proposed a high-throughput electrochemical detection platform called HiMEX, which could distinguish colorectal cancer patients from healthy volunteers with high sensitivity and specificity.…”

Section: Tumor-derived Evs As Biomarkers In Cancer Diagnosis and Treatmentmentioning

confidence: 99%

Tumor-Derived Extracellular Vesicles Regulate Cancer Progression in the Tumor Microenvironment

Bao

Huang

Chen

et al. 2022

Front. Mol. Biosci.

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Extracellular vesicles (EVs) are nanosized particles released by numerous kinds of cells, which are now increasingly considered as essential vehicles of cell-to-cell communication and biomarkers in disease diagnosis and treatment. They contain a variety of biomolecular components, including lipids, proteins and nucleic acids. These functional molecules can be transmitted between tumor cells and other stromal cells such as endothelial cells, fibroblasts and immune cells utilizing EVs. As a result, tumor-derived EVs can deliver molecules to remodel the tumor microenvironment, thereby influencing cancer progression. On the one hand, tumor-derived EVs reprogram functions of endothelial cells, promote cancer-associated fibroblasts transformation, induce resistance to therapy and inhibit the immune response to form a pro-tumorigenic environment. On the other hand, tumor-derived EVs stimulate the immune response to create an anti-tumoral environment. This article focuses on presenting a comprehensive and critical overview of the potential role of tumor-derived EVs-mediated communication in the tumor microenvironment.

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“…Analyzing EVs can enable clinicians to detect and monitor tumors in real time, while minimizing complications from specimen collection. 1 − 3 Profiling EV proteins, in particular, has been shown to achieve high diagnostic accuracy by targeting tumor-specific antigens, 4 − 6 identify tumor origins or subtypes based on the EV-protein signature, 7 − 10 and distinguish between indolent and high-risk lesions. 11 New assay formats have been developed to facilitate such EV-protein detection, many of which demonstrated high analytical sensitivity and throughput (see Table S1 for a comparison of methods).…”

Section: Introductionmentioning

confidence: 99%

Multielectrode Spectroscopy Enables Rapid and Sensitive Molecular Profiling of Extracellular Vesicles

et al. 2022

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Detecting protein markers in extracellular vesicles (EVs) is becoming a useful tool for basic research and clinical diagnoses. Most EV protein assays, however, require lengthy processes—conjugating affinity ligands onto sensing substrates and affixing EVs with additional labels to maximize signal generation. Here, we present an iPEX (impedance profiling of extracellular vesicles) system, an all-electrical strategy toward fast, multiplexed EV profiling. iPEX adopts one-step electropolymerization to rapidly functionalize sensor electrodes with antibodies; it then detects EV proteins in a label-free manner through impedance spectroscopy. The approach streamlines the entire EV assay, from sensor preparation to signal measurements. We achieved (i) fast immobilization of antibodies (<3 min) per electrode; (ii) high sensitivity (500 EVs/mL) without secondary labeling; and (iii) parallel detection (quadruple) in a single chip. A potential clinical utility was demonstrated by directly analyzing plasma samples from glioblastoma multiforme patients.

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An integrated magneto-electrochemical device for the rapid profiling of tumour extracellular vesicles from blood plasma

Cited by 111 publications

References 62 publications

Characterization and modulation of surface charges to enhance extracellular vesicle isolation in plasma

Characterization and modulation of surface charges to enhance extracellular vesicle isolation in plasma

Tumor-Derived Extracellular Vesicles Regulate Cancer Progression in the Tumor Microenvironment

Multielectrode Spectroscopy Enables Rapid and Sensitive Molecular Profiling of Extracellular Vesicles

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