EV-TRACK

Search > Results

You searched for: EV200076 (EV-TRACK ID)

Showing 1 - 5 of 5

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200076	1/5	microalgae	Dinoflagellate	(d)(U)C	Picciotto, Sabrina	2021	67%
Study summary Full title Isolation of extracellular vesicles from microalgae: towards the production of sustainable and natural nanocarriers of bioactive compounds All authors Sabrina Picciotto, Maria E. Barone, David Fierli, Anita Aranyos, Giorgia Adamo, Darja Božič, Daniele P. Romancino, Christopher Stanly, Rachel Parkes, Svenja Morsbach, Samuele Raccosta, Carolina Paganini, Antonella Cusimano, Vincenzo Martorana, Rosina Noto, Rita Carrotta, Fabio Librizzi, Umberto Capasso Palmiero, Pamela Santonicola, Ales Iglič, Meiyu Gai, Laura Corcuera, Annamaria Kisslinger, Elia Di Schiavi, Katharina Landfester, Giovanna L. Liguori, Veronika Kralj-Iglič, Paolo Arosio, Gabriella Pocsfalvi, Mauro Manno, Nicolas Touzet, Antonella Bongiovanni Journal Biomaterials science Abstract Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeuti (show more...)Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeutics, precision medicine and other biotechnology sectors. Novel bio-based nanotechnologies have recently arisen, which are based on the exploitation of extracellular vesicles (EVs). In this context, it has become essential to identify suitable organisms or cellular types to act as reliable sources of EVs and to develop their pilot- to large-scale production. The discovery of new biosources and the optimisation of related bioprocesses for the isolation and functionalisation of nano-delivery vehicles are fundamental to further develop therapeutic and biotechnological applications. Microalgae constitute sustainable sources of bioactive compounds with a range of sectorial applications including for example the formulation of health supplements, cosmetic products or food ingredients. In this study, we demonstrate that microalgae are promising producers of EVs. By analysing the nanosized extracellular nano-objects produced by eighteen microalgal species, we identified seven promising EV-producing strains belonging to distinct lineages, suggesting that the production of EVs in microalgae is an evolutionary conserved trait. Here we report the selection process and focus on one of this seven species, the glaucophyte Cyanophora paradoxa, which returned a protein yield in the small EV fraction of 1 μg of EV proteins per mg of dry weight of microalgal biomass (corresponding to 109 particles per mg of dried biomass) and EVs with a diameter of 130 nm (mode), as determined by the micro bicinchoninic acid assay, nanoparticle tracking and dynamic light scattering analyses. Moreover, the extracellular nanostructures isolated from the conditioned media of microalgae species returned positive immunoblot signals for some commonly used EV-biomarkers such as Alix, Enolase, HSP70, and β-actin. Overall, this work establishes a platform for the efficient production of EVs from a sustainable bioresource and highlights the potential of microalgal EVs as novel biogenic nanovehicles. (hide) EV-METRIC 67% (94th 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 (d)(U)C Protein markers EV: Alix/ HSP70/ beta-actin/ enolase non-EV: None Proteomics no Show all info Study aim Function/Biomarker/novel EV type Sample Species microalgae Sample Type Cell culture supernatant EV-producing cells Dinoflagellate EV-harvesting Medium Serum free medium Cell viability (%) 90 Cell count 1 mg dry weight biomass/ml Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type SW 28 Pelleting: speed (g) 118000 Wash: volume per pellet (ml) 32 Wash: time (min) 70 Wash: Rotor Type SW 28 Wash: speed (g) 118000 Characterization: Protein analysis Protein Concentration Method microBCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins beta-actin/ enolase/ HSP70/ Alix Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Size range/distribution Reported size (nm) 92 NTA Report type Size range/distribution Reported size (nm) 125 EV concentration Yes Particle yield number of particles per mg dry weight microalgal mass 6.00E+09 EM EM-type Scanning-EM Image type Close-up, Wide-field

	EV200076	2/5	microalgae	Diatom	(d)(U)C	Picciotto, Sabrina	2021	67%
Study summary Full title Isolation of extracellular vesicles from microalgae: towards the production of sustainable and natural nanocarriers of bioactive compounds All authors Sabrina Picciotto, Maria E. Barone, David Fierli, Anita Aranyos, Giorgia Adamo, Darja Božič, Daniele P. Romancino, Christopher Stanly, Rachel Parkes, Svenja Morsbach, Samuele Raccosta, Carolina Paganini, Antonella Cusimano, Vincenzo Martorana, Rosina Noto, Rita Carrotta, Fabio Librizzi, Umberto Capasso Palmiero, Pamela Santonicola, Ales Iglič, Meiyu Gai, Laura Corcuera, Annamaria Kisslinger, Elia Di Schiavi, Katharina Landfester, Giovanna L. Liguori, Veronika Kralj-Iglič, Paolo Arosio, Gabriella Pocsfalvi, Mauro Manno, Nicolas Touzet, Antonella Bongiovanni Journal Biomaterials science Abstract Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeuti (show more...)Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeutics, precision medicine and other biotechnology sectors. Novel bio-based nanotechnologies have recently arisen, which are based on the exploitation of extracellular vesicles (EVs). In this context, it has become essential to identify suitable organisms or cellular types to act as reliable sources of EVs and to develop their pilot- to large-scale production. The discovery of new biosources and the optimisation of related bioprocesses for the isolation and functionalisation of nano-delivery vehicles are fundamental to further develop therapeutic and biotechnological applications. Microalgae constitute sustainable sources of bioactive compounds with a range of sectorial applications including for example the formulation of health supplements, cosmetic products or food ingredients. In this study, we demonstrate that microalgae are promising producers of EVs. By analysing the nanosized extracellular nano-objects produced by eighteen microalgal species, we identified seven promising EV-producing strains belonging to distinct lineages, suggesting that the production of EVs in microalgae is an evolutionary conserved trait. Here we report the selection process and focus on one of this seven species, the glaucophyte Cyanophora paradoxa, which returned a protein yield in the small EV fraction of 1 μg of EV proteins per mg of dry weight of microalgal biomass (corresponding to 109 particles per mg of dried biomass) and EVs with a diameter of 130 nm (mode), as determined by the micro bicinchoninic acid assay, nanoparticle tracking and dynamic light scattering analyses. Moreover, the extracellular nanostructures isolated from the conditioned media of microalgae species returned positive immunoblot signals for some commonly used EV-biomarkers such as Alix, Enolase, HSP70, and β-actin. Overall, this work establishes a platform for the efficient production of EVs from a sustainable bioresource and highlights the potential of microalgal EVs as novel biogenic nanovehicles. (hide) EV-METRIC 67% (94th 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 (d)(U)C Protein markers EV: Alix/ HSP70/ enolase non-EV: None Proteomics no Show all info Study aim Function/Biomarker/novel EV type Sample Species microalgae Sample Type Cell culture supernatant EV-producing cells Diatom EV-harvesting Medium Serum free medium Cell viability (%) 90 Cell count 1 mg dry weight biomass/ml Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type SW 28 Pelleting: speed (g) 118000 Wash: volume per pellet (ml) 32 Wash: time (min) 70 Wash: Rotor Type SW 28 Wash: speed (g) 118000 Characterization: Protein analysis Protein Concentration Method microBCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins enolase/ HSP70/ Alix Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Size range/distribution Reported size (nm) 135 NTA Report type Size range/distribution Reported size (nm) 90 EV concentration Yes Particle yield number of particles per mg dry weight microalgal mass 2.40E+08 EM EM-type Scanning-EM Image type Close-up, Wide-field

	EV200076	3/5	microalgae	Glaucophyte	(d)(U)C	Picciotto, Sabrina	2021	67%
Study summary Full title Isolation of extracellular vesicles from microalgae: towards the production of sustainable and natural nanocarriers of bioactive compounds All authors Sabrina Picciotto, Maria E. Barone, David Fierli, Anita Aranyos, Giorgia Adamo, Darja Božič, Daniele P. Romancino, Christopher Stanly, Rachel Parkes, Svenja Morsbach, Samuele Raccosta, Carolina Paganini, Antonella Cusimano, Vincenzo Martorana, Rosina Noto, Rita Carrotta, Fabio Librizzi, Umberto Capasso Palmiero, Pamela Santonicola, Ales Iglič, Meiyu Gai, Laura Corcuera, Annamaria Kisslinger, Elia Di Schiavi, Katharina Landfester, Giovanna L. Liguori, Veronika Kralj-Iglič, Paolo Arosio, Gabriella Pocsfalvi, Mauro Manno, Nicolas Touzet, Antonella Bongiovanni Journal Biomaterials science Abstract Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeuti (show more...)Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeutics, precision medicine and other biotechnology sectors. Novel bio-based nanotechnologies have recently arisen, which are based on the exploitation of extracellular vesicles (EVs). In this context, it has become essential to identify suitable organisms or cellular types to act as reliable sources of EVs and to develop their pilot- to large-scale production. The discovery of new biosources and the optimisation of related bioprocesses for the isolation and functionalisation of nano-delivery vehicles are fundamental to further develop therapeutic and biotechnological applications. Microalgae constitute sustainable sources of bioactive compounds with a range of sectorial applications including for example the formulation of health supplements, cosmetic products or food ingredients. In this study, we demonstrate that microalgae are promising producers of EVs. By analysing the nanosized extracellular nano-objects produced by eighteen microalgal species, we identified seven promising EV-producing strains belonging to distinct lineages, suggesting that the production of EVs in microalgae is an evolutionary conserved trait. Here we report the selection process and focus on one of this seven species, the glaucophyte Cyanophora paradoxa, which returned a protein yield in the small EV fraction of 1 μg of EV proteins per mg of dry weight of microalgal biomass (corresponding to 109 particles per mg of dried biomass) and EVs with a diameter of 130 nm (mode), as determined by the micro bicinchoninic acid assay, nanoparticle tracking and dynamic light scattering analyses. Moreover, the extracellular nanostructures isolated from the conditioned media of microalgae species returned positive immunoblot signals for some commonly used EV-biomarkers such as Alix, Enolase, HSP70, and β-actin. Overall, this work establishes a platform for the efficient production of EVs from a sustainable bioresource and highlights the potential of microalgal EVs as novel biogenic nanovehicles. (hide) EV-METRIC 67% (94th 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 (d)(U)C Protein markers EV: Alix/ HSP70/ beta-actin/ enolase non-EV: None Proteomics no Show all info Study aim Function/Biomarker/novel EV type Sample Species microalgae Sample Type Cell culture supernatant EV-producing cells Glaucophyte EV-harvesting Medium Serum free medium Cell viability (%) 90 Cell count 1 mg dry weight biomass/ml Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type SW 28 Pelleting: speed (g) 118000 Wash: volume per pellet (ml) 32 Wash: time (min) 70 Wash: Rotor Type SW 28 Wash: speed (g) 118000 Characterization: Protein analysis Protein Concentration Method microBCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins Alix/ HSP70/ beta-actin/ enolase Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Size range/distribution Reported size (nm) 125 NTA Report type Size range/distribution Reported size (nm) 122 EV concentration Yes Particle yield number of particles per mg dry weight microalgal mass 2.00E+09 EM EM-type Scanning-EM Image type Close-up, Wide-field

	EV200076	4/5	microalgae	Haptophyte	(d)(U)C	Picciotto, Sabrina	2021	67%
Study summary Full title Isolation of extracellular vesicles from microalgae: towards the production of sustainable and natural nanocarriers of bioactive compounds All authors Sabrina Picciotto, Maria E. Barone, David Fierli, Anita Aranyos, Giorgia Adamo, Darja Božič, Daniele P. Romancino, Christopher Stanly, Rachel Parkes, Svenja Morsbach, Samuele Raccosta, Carolina Paganini, Antonella Cusimano, Vincenzo Martorana, Rosina Noto, Rita Carrotta, Fabio Librizzi, Umberto Capasso Palmiero, Pamela Santonicola, Ales Iglič, Meiyu Gai, Laura Corcuera, Annamaria Kisslinger, Elia Di Schiavi, Katharina Landfester, Giovanna L. Liguori, Veronika Kralj-Iglič, Paolo Arosio, Gabriella Pocsfalvi, Mauro Manno, Nicolas Touzet, Antonella Bongiovanni Journal Biomaterials science Abstract Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeuti (show more...)Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeutics, precision medicine and other biotechnology sectors. Novel bio-based nanotechnologies have recently arisen, which are based on the exploitation of extracellular vesicles (EVs). In this context, it has become essential to identify suitable organisms or cellular types to act as reliable sources of EVs and to develop their pilot- to large-scale production. The discovery of new biosources and the optimisation of related bioprocesses for the isolation and functionalisation of nano-delivery vehicles are fundamental to further develop therapeutic and biotechnological applications. Microalgae constitute sustainable sources of bioactive compounds with a range of sectorial applications including for example the formulation of health supplements, cosmetic products or food ingredients. In this study, we demonstrate that microalgae are promising producers of EVs. By analysing the nanosized extracellular nano-objects produced by eighteen microalgal species, we identified seven promising EV-producing strains belonging to distinct lineages, suggesting that the production of EVs in microalgae is an evolutionary conserved trait. Here we report the selection process and focus on one of this seven species, the glaucophyte Cyanophora paradoxa, which returned a protein yield in the small EV fraction of 1 μg of EV proteins per mg of dry weight of microalgal biomass (corresponding to 109 particles per mg of dried biomass) and EVs with a diameter of 130 nm (mode), as determined by the micro bicinchoninic acid assay, nanoparticle tracking and dynamic light scattering analyses. Moreover, the extracellular nanostructures isolated from the conditioned media of microalgae species returned positive immunoblot signals for some commonly used EV-biomarkers such as Alix, Enolase, HSP70, and β-actin. Overall, this work establishes a platform for the efficient production of EVs from a sustainable bioresource and highlights the potential of microalgal EVs as novel biogenic nanovehicles. (hide) EV-METRIC 67% (94th 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 (d)(U)C Protein markers EV: Alix/ HSP70/ enolase non-EV: None Proteomics no Show all info Study aim Function/Biomarker/novel EV type Sample Species microalgae Sample Type Cell culture supernatant EV-producing cells Haptophyte EV-harvesting Medium Serum free medium Cell viability (%) 90 Cell count 1 mg dry weight biomass/ml Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type SW 28 Pelleting: speed (g) 118000 Wash: volume per pellet (ml) 32 Wash: time (min) 70 Wash: Rotor Type SW 28 Wash: speed (g) 118000 Characterization: Protein analysis Protein Concentration Method microBCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins Alix/ enolase Not detected EV-associated proteins HSP70 Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Size range/distribution Reported size (nm) 87 NTA Report type Size range/distribution Reported size (nm) 165 EV concentration Yes Particle yield number of particles per mg dry weight microalgal mass 1.30E+08 EM EM-type Scanning-EM Image type Close-up, Wide-field

	EV200076	5/5	microalgae	Chlorophyte	(d)(U)C	Picciotto, Sabrina	2021	67%
Study summary Full title Isolation of extracellular vesicles from microalgae: towards the production of sustainable and natural nanocarriers of bioactive compounds All authors Sabrina Picciotto, Maria E. Barone, David Fierli, Anita Aranyos, Giorgia Adamo, Darja Božič, Daniele P. Romancino, Christopher Stanly, Rachel Parkes, Svenja Morsbach, Samuele Raccosta, Carolina Paganini, Antonella Cusimano, Vincenzo Martorana, Rosina Noto, Rita Carrotta, Fabio Librizzi, Umberto Capasso Palmiero, Pamela Santonicola, Ales Iglič, Meiyu Gai, Laura Corcuera, Annamaria Kisslinger, Elia Di Schiavi, Katharina Landfester, Giovanna L. Liguori, Veronika Kralj-Iglič, Paolo Arosio, Gabriella Pocsfalvi, Mauro Manno, Nicolas Touzet, Antonella Bongiovanni Journal Biomaterials science Abstract Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeuti (show more...)Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeutics, precision medicine and other biotechnology sectors. Novel bio-based nanotechnologies have recently arisen, which are based on the exploitation of extracellular vesicles (EVs). In this context, it has become essential to identify suitable organisms or cellular types to act as reliable sources of EVs and to develop their pilot- to large-scale production. The discovery of new biosources and the optimisation of related bioprocesses for the isolation and functionalisation of nano-delivery vehicles are fundamental to further develop therapeutic and biotechnological applications. Microalgae constitute sustainable sources of bioactive compounds with a range of sectorial applications including for example the formulation of health supplements, cosmetic products or food ingredients. In this study, we demonstrate that microalgae are promising producers of EVs. By analysing the nanosized extracellular nano-objects produced by eighteen microalgal species, we identified seven promising EV-producing strains belonging to distinct lineages, suggesting that the production of EVs in microalgae is an evolutionary conserved trait. Here we report the selection process and focus on one of this seven species, the glaucophyte Cyanophora paradoxa, which returned a protein yield in the small EV fraction of 1 μg of EV proteins per mg of dry weight of microalgal biomass (corresponding to 109 particles per mg of dried biomass) and EVs with a diameter of 130 nm (mode), as determined by the micro bicinchoninic acid assay, nanoparticle tracking and dynamic light scattering analyses. Moreover, the extracellular nanostructures isolated from the conditioned media of microalgae species returned positive immunoblot signals for some commonly used EV-biomarkers such as Alix, Enolase, HSP70, and β-actin. Overall, this work establishes a platform for the efficient production of EVs from a sustainable bioresource and highlights the potential of microalgal EVs as novel biogenic nanovehicles. (hide) EV-METRIC 67% (94th 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 (d)(U)C Protein markers EV: Alix/ beta-actin/ enolase non-EV: None Proteomics no Show all info Study aim Function/Biomarker/novel EV type Sample Species microalgae Sample Type Cell culture supernatant EV-producing cells Chlorophyte EV-harvesting Medium Serum free medium Cell viability (%) 90 Cell count 1 mg dry weight biomass/ml Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type SW 28 Pelleting: speed (g) 118000 Wash: volume per pellet (ml) 32 Wash: time (min) 70 Wash: Rotor Type SW 28 Wash: speed (g) 118000 Characterization: Protein analysis Protein Concentration Method microBCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins beta-actin/ enolase/ Alix Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Size range/distribution Reported size (nm) 75 NTA Report type Size range/distribution Reported size (nm) 137 EV concentration Yes Particle yield number of particles per mg dry weight microalgal mass 2.60E+08 EM EM-type Scanning-EM Image type Close-up, Wide-field

				1 - 5 of 5

EV-TRACK ID	EV200076
species	microalgae
sample type	Cell culture
cell type	Dinoflagellate	Diatom	Glaucophyte	Haptophyte	Chlorophyte
condition	Control condition	Control condition	Control condition	Control condition	Control condition
separation protocol	(d)(U)C	(d)(U)C	(d)(U)C	(d)(U)C	(d)(U)C
Exp. nr.	1	2	3	4	5
EV-METRIC %	67	67	67	67	67