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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV240006	3/4	Rattus norvegicus	INS-1	(d)(U)C Total Exosome Isolation lipid-based affinity capture	Weerakkody, Jonathan S.	2024	50%
Study summary Full title Photosensitive Nanoprobes for Rapid High Purity Isolation and Size-Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations All authors Jonathan S. Weerakkody, Tiffany Tseng, Mackenzie Topper, Sikha Thoduvayil, Abhijith Radhakrishnan, Frederic Pincet, Themis R. Kyriakides, Roshan W. Gunasekara, Sathish Ramakrishnan Journal Abstract The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as (show more...)The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as exosomes, secretory, and synthetic vesicles, lies in the absence of a unified approach that seamlessly delivers high purity, yield, and scalability for large-scale applications. To address this gap, an innovative method is developed that utilizes photosensitive lipid nanoprobes for the efficient isolation of vesicles and sorting them into subpopulations based on size. The photosensitive component in the probe undergoes cleavage upon exposure to light, facilitating the release of vesicles in their near-native form. The method demonstrates a superior ability in isolating high purity extracellular vesicles from complex biological media and separating them into size-based subpopulations within 1 h, achieving more efficiency and purity than ultracentrifugation. Furthermore, this method's cost-effectiveness and rapid enrichment of the vesicles align with demands for large-scale isolation and downstream analyses of nucleic acids and proteins. The method opens new avenues in exploring, analyzing, and utilizing synthetic and extracellular vesicle subpopulations in various biomedical applications, including diagnostics, therapeutic delivery, and biomarker discovery. (hide) EV-METRIC 50% (87th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles small extracellular vesicles Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Total Exosome Isolation lipid-based affinity capture Protein markers EV: CD9/ CD81 non-EV: None Proteomics no Show all info Study aim New methodological development/Technical analysis comparing/optimizing EV-related methods Sample Species Rattus norvegicus Sample Type Cell culture supernatant EV-producing cells INS-1 EV-harvesting Medium EV-depleted medium Preparation of EDS Commercial EDS Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Pelleting performed Yes Pelleting: rotor type SW 55 Ti Pelleting: speed (g) 12500 Commercial kit Total Exosome Isolation Other Name other separation method Total Exosome Isolation Other Name other separation method lipid-based affinity capture EV-subtype Distinction between multiple subtypes Size Used subtypes 30-200 Characterization: Protein analysis Protein Concentration Method Not determined Detected EV-associated proteins CD9/ CD81 Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 104 NTA Report type Mean Reported size (nm) 126 EV concentration Yes Particle yield particles per milliliter of starting sample: 1.50E+07 EM EM-type Transmission-EM Image type Close-up, Wide-field Other particle analysis name(1) dSTORM single molecule localization microscopy Report type Mean Report size 89 EV-concentration No Extra information This paper was to validate the efficacy of photosensitive lipid nanoprobe for the isolation and size selective enrichment of native extracellular vesicles

	EV240006	4/4	Mus musculus	Neuro-2a (N2a) CCL-131	(d)(U)C Total Exosome Isolation lipid-based affinity capture	Weerakkody, Jonathan S.	2024	50%
Study summary Full title Photosensitive Nanoprobes for Rapid High Purity Isolation and Size-Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations All authors Jonathan S. Weerakkody, Tiffany Tseng, Mackenzie Topper, Sikha Thoduvayil, Abhijith Radhakrishnan, Frederic Pincet, Themis R. Kyriakides, Roshan W. Gunasekara, Sathish Ramakrishnan Journal Abstract The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as (show more...)The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as exosomes, secretory, and synthetic vesicles, lies in the absence of a unified approach that seamlessly delivers high purity, yield, and scalability for large-scale applications. To address this gap, an innovative method is developed that utilizes photosensitive lipid nanoprobes for the efficient isolation of vesicles and sorting them into subpopulations based on size. The photosensitive component in the probe undergoes cleavage upon exposure to light, facilitating the release of vesicles in their near-native form. The method demonstrates a superior ability in isolating high purity extracellular vesicles from complex biological media and separating them into size-based subpopulations within 1 h, achieving more efficiency and purity than ultracentrifugation. Furthermore, this method's cost-effectiveness and rapid enrichment of the vesicles align with demands for large-scale isolation and downstream analyses of nucleic acids and proteins. The method opens new avenues in exploring, analyzing, and utilizing synthetic and extracellular vesicle subpopulations in various biomedical applications, including diagnostics, therapeutic delivery, and biomarker discovery. (hide) EV-METRIC 50% (87th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles small extracellular vesicles Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Total Exosome Isolation lipid-based affinity capture Protein markers EV: CD9/ CD81 non-EV: None Proteomics no Show all info Study aim New methodological development/Technical analysis comparing/optimizing EV-related methods Sample Species Mus musculus Sample Type Cell culture supernatant EV-producing cells Neuro-2a (N2a) CCL-131 EV-harvesting Medium EV-depleted medium Preparation of EDS Commercial EDS Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Pelleting performed Yes Pelleting: rotor type SW 55 Ti Pelleting: speed (g) 12500 Commercial kit Total Exosome Isolation Other Name other separation method Total Exosome Isolation Other Name other separation method lipid-based affinity capture EV-subtype Distinction between multiple subtypes Size Used subtypes 150-350 Characterization: Protein analysis Protein Concentration Method Not determined Detected EV-associated proteins CD9/ CD81 Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 105 NTA Report type Mean Reported size (nm) 122 EV concentration Yes Particle yield particles per milliliter of starting sample: 3.00E+07 EM EM-type Transmission-EM Image type Close-up, Wide-field Other particle analysis name(1) dSTORM single molecule localization microscopy Report type Mean Report size 52 EV-concentration No Extra information This paper was to validate the efficacy of photosensitive lipid nanoprobe for the isolation and size selective enrichment of native extracellular vesicles

	EV240006	1/4	Mus musculus	Neuro-2a (N2a) CCL-131	(d)(U)C Total Exosome Isolation lipid-based affinity capture	Weerakkody, Jonathan S.	2024	44%
Study summary Full title Photosensitive Nanoprobes for Rapid High Purity Isolation and Size-Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations All authors Jonathan S. Weerakkody, Tiffany Tseng, Mackenzie Topper, Sikha Thoduvayil, Abhijith Radhakrishnan, Frederic Pincet, Themis R. Kyriakides, Roshan W. Gunasekara, Sathish Ramakrishnan Journal Abstract The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as (show more...)The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as exosomes, secretory, and synthetic vesicles, lies in the absence of a unified approach that seamlessly delivers high purity, yield, and scalability for large-scale applications. To address this gap, an innovative method is developed that utilizes photosensitive lipid nanoprobes for the efficient isolation of vesicles and sorting them into subpopulations based on size. The photosensitive component in the probe undergoes cleavage upon exposure to light, facilitating the release of vesicles in their near-native form. The method demonstrates a superior ability in isolating high purity extracellular vesicles from complex biological media and separating them into size-based subpopulations within 1 h, achieving more efficiency and purity than ultracentrifugation. Furthermore, this method's cost-effectiveness and rapid enrichment of the vesicles align with demands for large-scale isolation and downstream analyses of nucleic acids and proteins. The method opens new avenues in exploring, analyzing, and utilizing synthetic and extracellular vesicle subpopulations in various biomedical applications, including diagnostics, therapeutic delivery, and biomarker discovery. (hide) EV-METRIC 44% (84th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles small extracellular vesicles Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Total Exosome Isolation lipid-based affinity capture Protein markers EV: CD9/ CD63/ CD81/ Syt1 non-EV: None Proteomics no Show all info Study aim New methodological development/Technical analysis comparing/optimizing EV-related methods Sample Species Mus musculus Sample Type Cell culture supernatant EV-producing cells Neuro-2a (N2a) CCL-131 EV-harvesting Medium EV-depleted medium Preparation of EDS Commercial EDS Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Pelleting performed Yes Pelleting: rotor type SW 55 Ti Pelleting: speed (g) 12500 Commercial kit Total Exosome Isolation Other Name other separation method Total Exosome Isolation Other Name other separation method lipid-based affinity capture EV-subtype Distinction between multiple subtypes Size Used subtypes 30-150 Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins CD9/ CD63/ CD81/ Syt1 Detected EV-associated proteins CD9/ CD63/ Syt1 Characterization: RNA analysis RNA analysis Type RNA-sequencing Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 100-150 Extra information This paper was to validate the efficacy of photosensitive lipid nanoprobe for the isolation and size selective enrichment of native extracellular vesicles

	EV240006	2/4	Mus musculus	Primary bone marrow-derived macrophage (BMDM) cells from hind limbs in C57BL/6J wild type mice	(d)(U)C Total Exosome Isolation lipid-based affinity capture	Weerakkody, Jonathan S.	2024	44%
Study summary Full title Photosensitive Nanoprobes for Rapid High Purity Isolation and Size-Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations All authors Jonathan S. Weerakkody, Tiffany Tseng, Mackenzie Topper, Sikha Thoduvayil, Abhijith Radhakrishnan, Frederic Pincet, Themis R. Kyriakides, Roshan W. Gunasekara, Sathish Ramakrishnan Journal Abstract The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as (show more...)The biggest challenge in current isolation methods for lipid bilayer-encapsulated vesicles, such as exosomes, secretory, and synthetic vesicles, lies in the absence of a unified approach that seamlessly delivers high purity, yield, and scalability for large-scale applications. To address this gap, an innovative method is developed that utilizes photosensitive lipid nanoprobes for the efficient isolation of vesicles and sorting them into subpopulations based on size. The photosensitive component in the probe undergoes cleavage upon exposure to light, facilitating the release of vesicles in their near-native form. The method demonstrates a superior ability in isolating high purity extracellular vesicles from complex biological media and separating them into size-based subpopulations within 1 h, achieving more efficiency and purity than ultracentrifugation. Furthermore, this method's cost-effectiveness and rapid enrichment of the vesicles align with demands for large-scale isolation and downstream analyses of nucleic acids and proteins. The method opens new avenues in exploring, analyzing, and utilizing synthetic and extracellular vesicle subpopulations in various biomedical applications, including diagnostics, therapeutic delivery, and biomarker discovery. (hide) EV-METRIC 44% (84th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Total Exosome Isolation lipid-based affinity capture Protein markers EV: CD9/ CD63/ CD81/ Syt1 non-EV: None Proteomics no Show all info Study aim New methodological development/Technical analysis comparing/optimizing EV-related methods Sample Species Mus musculus Sample Type Cell culture supernatant EV-producing cells Primary bone marrow-derived macrophage (BMDM) cells from hind limbs in C57BL/6J wild type mice EV-harvesting Medium EV-depleted medium Preparation of EDS Commercial EDS Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Pelleting performed Yes Pelleting: rotor type SW 55 Ti Pelleting: speed (g) 12500 Commercial kit Total Exosome Isolation Other Name other separation method Total Exosome Isolation Other Name other separation method lipid-based affinity capture EV-subtype Distinction between multiple subtypes Size Used subtypes 30-200 Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins CD9/ CD63/ CD81/ Syt1 Detected EV-associated proteins CD9/ CD63/ Syt1 Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 220-350 Extra information This paper was to validate the efficacy of photosensitive lipid nanoprobe for the isolation and size selective enrichment of native extracellular vesicles

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EV-TRACK ID	EV240006
species	Rattus norvegicus	Mus musculus	Mus musculus	Mus musculus
sample type	Cell culture	Cell culture	Cell culture	Cell culture
cell type	INS-1	Neuro-2a (N2a) CCL-131	Neuro-2a (N2a) CCL-131	Primary bone marrow-derived macrophage (BMDM) cells from hind limbs in C57BL/6J wild type mice
condition	Control condition	Control condition	Control condition	Control condition
separation protocol	dUC/ Total Exosome Isolation/ lipid-based affinity capture	dUC/ Total Exosome Isolation/ lipid-based affinity capture	dUC/ Total Exosome Isolation/ lipid-based affinity capture	dUC/ Total Exosome Isolation/ lipid-based affinity capture
EV subtype	30-200	150-350	30-150	30-200
vesicle related term	small EVs	small EVs	small EVs	EV
Exp. nr.	3	4	1	2
EV-METRIC %	50	50	44	44