András Saftics | Budapest University of Technology and Economics (original) (raw)
Papers by András Saftics
Journal of the American College of Cardiology, Apr 1, 2024
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Journal of biological chemistry/The Journal of biological chemistry, Mar 1, 2024
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Journal of extracellular vesicles, Feb 1, 2024
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Cancers, Jan 27, 2024
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Circulation Research, Aug 3, 2023
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Extracellular vesicle, Nov 30, 2023
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arXiv (Cornell University), Mar 27, 2020
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Journal of extracellular vesicles, Jul 1, 2023
Extracellular vesicles (EVs) and their cargo constitute novel biomarkers. EV subpopulations have ... more Extracellular vesicles (EVs) and their cargo constitute novel biomarkers. EV subpopulations have been defined not only by abundant tetraspanins (e.g., CD9, CD63 and CD81) but also by specific markers derived from their source cells. However, it remains a challenge to robustly isolate and characterize EV subpopulations. Here, we combined affinity isolation with super‐resolution imaging to comprehensively assess EV subpopulations from human plasma. Our Single Extracellular VEsicle Nanoscopy (SEVEN) assay successfully quantified the number of affinity‐isolated EVs, their size, shape, molecular tetraspanin content, and heterogeneity. The number of detected tetraspanin‐enriched EVs positively correlated with sample dilution in a 64‐fold range (for SEC‐enriched plasma) and a 50‐fold range (for crude plasma). Importantly, SEVEN robustly detected EVs from as little as ∼0.1 μL of crude plasma. We further characterized the size, shape and molecular tetraspanin content (with corresponding heterogeneities) for CD9‐, CD63‐ and CD81‐enriched EV subpopulations. Finally, we assessed EVs from the plasma of four pancreatic ductal adenocarcinoma patients with resectable disease. Compared to healthy plasma, CD9‐enriched EVs from patients were smaller while IGF1R‐enriched EVs from patients were larger, rounder and contained more tetraspanin molecules, suggestive of a unique pancreatic cancer‐enriched EV subpopulation. This study provides the method validation and demonstrates that SEVEN could be advanced into a platform for characterizing both disease‐associated and organ‐associated EV subpopulations.
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bioRxiv (Cold Spring Harbor Laboratory), May 29, 2023
ABSTRACTExosomes are small extracellular vesicles (sEVs) of ∼30-150 nm in diameter that have the ... more ABSTRACTExosomes are small extracellular vesicles (sEVs) of ∼30-150 nm in diameter that have the same topology as the cell, are enriched in selected exosome cargo proteins, and play important roles in health and disease. To address large unanswered questions regarding exosome biologyin vivo, we created theexomap1transgenic mouse model. In response to Cre recombinase,exomap1mice express HsCD81mNG, a fusion protein between human CD81, the most highly enriched exosome protein yet described, and the bright green fluorescent protein mNeonGreen. As expected, cell type-specific expression of Cre induced the cell type-specific expression of HsCD81mNG in diverse cell types, correctly localized HsCD81mNG to the plasma membrane, and selectively loaded HsCD81mNG into secreted vesicles that have the size (∼80 nm), topology (outside out), and content (presence of mouse exosome markers) of exosomes. Furthermore, mouse cells expressing HsCD81mNG released HsCD81mNG-marked exosomes into blood and other biofluids. Using high-resolution, single-exosome analysis by quantitative single molecule localization microscopy, we show here that that hepatocytes contribute ∼15% of the blood exosome population whereas neurons contribute <1% of blood exosomes. These estimates of cell type-specific contributions to blood EV population are consistent with the porosity of liver sinusoidal endothelial cells to particles of ∼50-300 nm in diameter, as well as with the impermeability of blood-brain and blood-neuron barriers to particles >5 nm in size. Taken together, these results establish theexomap1mouse as a useful tool forin vivostudies of exosome biology, and for mapping cell type-specific contributions to biofluid exosome populations. In addition, our data confirm that CD81 is a highly-specific marker for exosomes and is not enriched in the larger microvesicle class of EVs.
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Scientific Reports, Jan 9, 2020
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Scientific Reports, Feb 24, 2020
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Analyst, 2020
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Applied Surface Science, 2021
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Langmuir, Jan 17, 2019
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Biosensors, Jan 20, 2022
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Applied Surface Science, Sep 1, 2013
ABSTRACT Novel biosensors made of polymers may offer advantages over conventional technology such... more ABSTRACT Novel biosensors made of polymers may offer advantages over conventional technology such as possibility of mass production and tunability of the material properties. With the ongoing work on the polymer photonic chip fabrication in our project, simple model samples were tested parallel for future immobilization and accessing conditions for applications in typical aqueous buffers. The model samples consist of a thin, high refractive index polyimide film on top of TEOS on Si wafer. These model samples were measured by in situ spectroscopic ellipsometry using different aqueous buffers. The experiments revealed a high drift in aqueous solutions; the drift in the ellipsometric parameters (delta, psi) can be evaluated and presented as changes in thickness and refractive index of the polyimide layer. The first molecular layer of immobilization is based on polyethyleneimine (PEI). The signal for the PEI adsorption was detected on a stable baseline, only after a long conditioning. The stability of polyimide films in aqueous buffer solutions should be improved toward the real biosensor application. Preliminary results are shown on the possibilities to protect the polyimide. Optical Waveguide Lightmode Spectroscopy (OWLS) has been used to demonstrate the shielding effect of the thin TiO2 adlayer in biosensor applications.
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Scientific Reports, Dec 17, 2020
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Colloids and interfaces, Feb 14, 2022
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Cancers, Jun 4, 2022
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Journal of the American College of Cardiology, Apr 1, 2024
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Journal of biological chemistry/The Journal of biological chemistry, Mar 1, 2024
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Journal of extracellular vesicles, Feb 1, 2024
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Cancers, Jan 27, 2024
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Circulation Research, Aug 3, 2023
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Extracellular vesicle, Nov 30, 2023
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arXiv (Cornell University), Mar 27, 2020
Bookmarks Related papers MentionsView impact
Journal of extracellular vesicles, Jul 1, 2023
Extracellular vesicles (EVs) and their cargo constitute novel biomarkers. EV subpopulations have ... more Extracellular vesicles (EVs) and their cargo constitute novel biomarkers. EV subpopulations have been defined not only by abundant tetraspanins (e.g., CD9, CD63 and CD81) but also by specific markers derived from their source cells. However, it remains a challenge to robustly isolate and characterize EV subpopulations. Here, we combined affinity isolation with super‐resolution imaging to comprehensively assess EV subpopulations from human plasma. Our Single Extracellular VEsicle Nanoscopy (SEVEN) assay successfully quantified the number of affinity‐isolated EVs, their size, shape, molecular tetraspanin content, and heterogeneity. The number of detected tetraspanin‐enriched EVs positively correlated with sample dilution in a 64‐fold range (for SEC‐enriched plasma) and a 50‐fold range (for crude plasma). Importantly, SEVEN robustly detected EVs from as little as ∼0.1 μL of crude plasma. We further characterized the size, shape and molecular tetraspanin content (with corresponding heterogeneities) for CD9‐, CD63‐ and CD81‐enriched EV subpopulations. Finally, we assessed EVs from the plasma of four pancreatic ductal adenocarcinoma patients with resectable disease. Compared to healthy plasma, CD9‐enriched EVs from patients were smaller while IGF1R‐enriched EVs from patients were larger, rounder and contained more tetraspanin molecules, suggestive of a unique pancreatic cancer‐enriched EV subpopulation. This study provides the method validation and demonstrates that SEVEN could be advanced into a platform for characterizing both disease‐associated and organ‐associated EV subpopulations.
Bookmarks Related papers MentionsView impact
bioRxiv (Cold Spring Harbor Laboratory), May 29, 2023
ABSTRACTExosomes are small extracellular vesicles (sEVs) of ∼30-150 nm in diameter that have the ... more ABSTRACTExosomes are small extracellular vesicles (sEVs) of ∼30-150 nm in diameter that have the same topology as the cell, are enriched in selected exosome cargo proteins, and play important roles in health and disease. To address large unanswered questions regarding exosome biologyin vivo, we created theexomap1transgenic mouse model. In response to Cre recombinase,exomap1mice express HsCD81mNG, a fusion protein between human CD81, the most highly enriched exosome protein yet described, and the bright green fluorescent protein mNeonGreen. As expected, cell type-specific expression of Cre induced the cell type-specific expression of HsCD81mNG in diverse cell types, correctly localized HsCD81mNG to the plasma membrane, and selectively loaded HsCD81mNG into secreted vesicles that have the size (∼80 nm), topology (outside out), and content (presence of mouse exosome markers) of exosomes. Furthermore, mouse cells expressing HsCD81mNG released HsCD81mNG-marked exosomes into blood and other biofluids. Using high-resolution, single-exosome analysis by quantitative single molecule localization microscopy, we show here that that hepatocytes contribute ∼15% of the blood exosome population whereas neurons contribute <1% of blood exosomes. These estimates of cell type-specific contributions to blood EV population are consistent with the porosity of liver sinusoidal endothelial cells to particles of ∼50-300 nm in diameter, as well as with the impermeability of blood-brain and blood-neuron barriers to particles >5 nm in size. Taken together, these results establish theexomap1mouse as a useful tool forin vivostudies of exosome biology, and for mapping cell type-specific contributions to biofluid exosome populations. In addition, our data confirm that CD81 is a highly-specific marker for exosomes and is not enriched in the larger microvesicle class of EVs.
Bookmarks Related papers MentionsView impact
Scientific Reports, Jan 9, 2020
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Scientific Reports, Feb 24, 2020
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Bookmarks Related papers MentionsView impact
Analyst, 2020
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Applied Surface Science, 2021
Bookmarks Related papers MentionsView impact
Langmuir, Jan 17, 2019
Bookmarks Related papers MentionsView impact
Biosensors, Jan 20, 2022
Bookmarks Related papers MentionsView impact
Applied Surface Science, Sep 1, 2013
ABSTRACT Novel biosensors made of polymers may offer advantages over conventional technology such... more ABSTRACT Novel biosensors made of polymers may offer advantages over conventional technology such as possibility of mass production and tunability of the material properties. With the ongoing work on the polymer photonic chip fabrication in our project, simple model samples were tested parallel for future immobilization and accessing conditions for applications in typical aqueous buffers. The model samples consist of a thin, high refractive index polyimide film on top of TEOS on Si wafer. These model samples were measured by in situ spectroscopic ellipsometry using different aqueous buffers. The experiments revealed a high drift in aqueous solutions; the drift in the ellipsometric parameters (delta, psi) can be evaluated and presented as changes in thickness and refractive index of the polyimide layer. The first molecular layer of immobilization is based on polyethyleneimine (PEI). The signal for the PEI adsorption was detected on a stable baseline, only after a long conditioning. The stability of polyimide films in aqueous buffer solutions should be improved toward the real biosensor application. Preliminary results are shown on the possibilities to protect the polyimide. Optical Waveguide Lightmode Spectroscopy (OWLS) has been used to demonstrate the shielding effect of the thin TiO2 adlayer in biosensor applications.
Bookmarks Related papers MentionsView impact
Scientific Reports, Dec 17, 2020
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Colloids and interfaces, Feb 14, 2022
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Cancers, Jun 4, 2022
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