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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230025	1/5	Homo sapiens	Milk	(d)(U)C qEV Filtration	Turner NP	2023	67%
Study summary Full title Omics Analysis of Extracellular Vesicles Recovered from Infant Formula Products and Milk: Towards Personalized Infant Nutrition. All authors Turner NP, Abeysinghe P, Sadowski P, Mitchell MD Journal Mol Nutr Food Res Abstract Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritiona (show more...)Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritional needs of the developing infant. Extracellular vesicles (EVs) in human (HM) and cow milk (CM) contain molecular cargo such as proteins and micro(mi)RNAs that serve as functional messengers between cells and may be of importance to infant health. Most IF is derived from a CM protein base, however differences between HM and CM EV molecular cargo have not been extensively studied. (hide) EV-METRIC 67% (81st 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 Milk 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 qEV Filtration Protein markers EV: CD9/ CD81/ Flotillin-1/ TSG101/ Syn-1/ GAPDH non-EV: Calnexin Proteomics yes Show all info Study aim Identification of content (omics approaches) Sample Species Homo sapiens Sample Type Milk Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type Type 50.2 Ti Pelleting: speed (g) 100,000 Filtration steps 0.2 or 0.22 µm Commercial kit qEV Other Name other separation method qEV Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) per milliliter of starting sample Western Blot Detected EV-associated proteins Flotillin-1/ TSG101/ Syntenin-1/ GAPDH/ CD81 Not detected contaminants Calnexin Proteomics database PRIDE Not detected contaminants Albumin Characterization: RNA analysis RNA analysis Type RNA-sequencing Database Vesiclepedia Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 125 EV concentration Yes Particle yield particles per milliliter of starting sample: 6.87E+08 EM EM-type Transmission-EM Image type Close-up, Wide-field

	EV230025	2/5	Bos taurus	Milk	(d)(U)C qEV Filtration	Turner NP	2023	67%
Study summary Full title Omics Analysis of Extracellular Vesicles Recovered from Infant Formula Products and Milk: Towards Personalized Infant Nutrition. All authors Turner NP, Abeysinghe P, Sadowski P, Mitchell MD Journal Mol Nutr Food Res Abstract Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritiona (show more...)Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritional needs of the developing infant. Extracellular vesicles (EVs) in human (HM) and cow milk (CM) contain molecular cargo such as proteins and micro(mi)RNAs that serve as functional messengers between cells and may be of importance to infant health. Most IF is derived from a CM protein base, however differences between HM and CM EV molecular cargo have not been extensively studied. (hide) EV-METRIC 67% (81st 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 Milk 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 qEV Filtration Protein markers EV: CD9/ CD81/ Flotillin-1/ TSG101/ Syn-1/ GAPDH non-EV: Albumin/ Calnexin Proteomics yes Show all info Study aim Identification of content (omics approaches) Sample Species Bos taurus Sample Type Milk Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type Type 50.2 Ti Pelleting: speed (g) 100,000 Filtration steps 0.2 or 0.22 µm Commercial kit qEV Other Name other separation method qEV Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) per milliliter of starting sample Western Blot Detected EV-associated proteins CD9/ CD81/ Flotillin-1/ TSG101/ Syntenin-1/ GAPDH Not detected contaminants Albumin/ Calnexin Proteomics database PRIDE Not detected contaminants Albumin Characterization: RNA analysis RNA analysis Type RNA-sequencing Database Vesiclepedia Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 100 EV concentration Yes Particle yield particles per milliliter of starting sample: 2.46E+10 EM EM-type Transmission-EM Image type Close-up, Wide-field

	EV230025	3/5	Bos taurus	Infant formula	(d)(U)C qEV Filtration	Turner NP	2023	67%
Study summary Full title Omics Analysis of Extracellular Vesicles Recovered from Infant Formula Products and Milk: Towards Personalized Infant Nutrition. All authors Turner NP, Abeysinghe P, Sadowski P, Mitchell MD Journal Mol Nutr Food Res Abstract Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritiona (show more...)Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritional needs of the developing infant. Extracellular vesicles (EVs) in human (HM) and cow milk (CM) contain molecular cargo such as proteins and micro(mi)RNAs that serve as functional messengers between cells and may be of importance to infant health. Most IF is derived from a CM protein base, however differences between HM and CM EV molecular cargo have not been extensively studied. (hide) EV-METRIC 67% (50th 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 Infant formula Sample origin IF 0-6 months 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 qEV Filtration Protein markers EV: CD9/ CD81/ Flotillin-1/ TSG101/ GAPDH non-EV: Albumin Proteomics yes Show all info Study aim Identification of content (omics approaches) Sample Species Bos taurus Sample Type Infant formula Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type Type 50.2 Ti Pelleting: speed (g) 100,000 Filtration steps 0.2 or 0.22 µm Commercial kit qEV Other Name other separation method qEV Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) per milliliter of starting sample Western Blot Detected EV-associated proteins CD9/ CD81/ TSG101/ Flotillin-1 Not detected EV-associated proteins GAPDH Detected contaminants Albumin Proteomics database PRIDE Detected contaminants Albumin Characterization: RNA analysis RNA analysis Type RNA-sequencing Database Vesiclepedia Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 105 EV concentration Yes Particle yield particles per milliliter of starting sample: 4.02E+09 EM EM-type Transmission-EM Image type Close-up, Wide-field

	EV230025	4/5	Bos taurus	Infant formula	(d)(U)C qEV Filtration	Turner NP	2023	67%
Study summary Full title Omics Analysis of Extracellular Vesicles Recovered from Infant Formula Products and Milk: Towards Personalized Infant Nutrition. All authors Turner NP, Abeysinghe P, Sadowski P, Mitchell MD Journal Mol Nutr Food Res Abstract Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritiona (show more...)Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritional needs of the developing infant. Extracellular vesicles (EVs) in human (HM) and cow milk (CM) contain molecular cargo such as proteins and micro(mi)RNAs that serve as functional messengers between cells and may be of importance to infant health. Most IF is derived from a CM protein base, however differences between HM and CM EV molecular cargo have not been extensively studied. (hide) EV-METRIC 67% (50th 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 Infant formula Sample origin IF 6-12 months 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 qEV Filtration Protein markers EV: CD9/ CD81/ Flotillin-1/ TSG101/ GAPDH non-EV: Albumin Proteomics yes Show all info Study aim Identification of content (omics approaches) Sample Species Bos taurus Sample Type Infant formula Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type Type 50.2 Ti Pelleting: speed (g) 100,000 Filtration steps 0.2 or 0.22 µm Commercial kit qEV Other Name other separation method qEV Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) per milliliter of starting sample Western Blot Detected EV-associated proteins CD9/ CD81/ TSG101/ Flotillin-1 Not detected EV-associated proteins GAPDH Detected contaminants Albumin Proteomics database PRIDE Detected contaminants Albumin Characterization: RNA analysis RNA analysis Type RNA-sequencing Database Vesiclepedia Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 100 EV concentration Yes Particle yield particles per milliliter of starting sample: 6.19E+09 EM EM-type Transmission-EM Image type Close-up, Wide-field

	EV230025	5/5	Bos taurus	Infant formula	(d)(U)C qEV Filtration	Turner NP	2023	67%
Study summary Full title Omics Analysis of Extracellular Vesicles Recovered from Infant Formula Products and Milk: Towards Personalized Infant Nutrition. All authors Turner NP, Abeysinghe P, Sadowski P, Mitchell MD Journal Mol Nutr Food Res Abstract Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritiona (show more...)Milk and milk products such as infant formula (IF) play a fundamental role in serving the nutritional needs of the developing infant. Extracellular vesicles (EVs) in human (HM) and cow milk (CM) contain molecular cargo such as proteins and micro(mi)RNAs that serve as functional messengers between cells and may be of importance to infant health. Most IF is derived from a CM protein base, however differences between HM and CM EV molecular cargo have not been extensively studied. (hide) EV-METRIC 67% (50th 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 Infant formula Sample origin IF 1 year+ 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 qEV Filtration Protein markers EV: CD9/ CD81/ Flotillin-1/ TSG101/ GAPDH non-EV: Albumin Proteomics yes Show all info Study aim Identification of content (omics approaches) Sample Species Bos taurus Sample Type Infant formula Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type Type 50.2 Ti Pelleting: speed (g) 100,000 Filtration steps 0.2 or 0.22 µm Commercial kit qEV Other Name other separation method qEV Characterization: Protein analysis Protein Concentration Method microBCA Protein Yield (µg) per milliliter of starting sample Western Blot Detected EV-associated proteins CD9/ CD81/ TSG101/ Flotillin-1 Not detected EV-associated proteins GAPDH Detected contaminants Albumin Proteomics database PRIDE Detected contaminants Albumin Characterization: RNA analysis RNA analysis Type RNA-sequencing Database Vesiclepedia Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 95 EV concentration Yes Particle yield particles per milliliter of starting sample: 1.95E+09 EM EM-type Transmission-EM Image type Close-up, Wide-field

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EV-TRACK ID	EV230025
species	Homo sapiens	Bos taurus	Bos taurus	Bos taurus	Bos taurus
sample type	Milk	Milk	Infant formula	Infant formula	Infant formula
condition	Control condition	Control condition	IF 0-6 months	IF 6-12 months	IF 1 year+
separation protocol	dUC/ qEV/ Filtration	dUC/ qEV/ Filtration	dUC/ qEV/ Filtration	dUC/ qEV/ Filtration	dUC/ qEV/ Filtration
Exp. nr.	1	2	3	4	5
EV-METRIC %	67	67	67	67	67