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You searched for: EV200076 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, separation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Separation protocol
  • Gives a short, non-chronological overview of the different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
Details EV-TRACK ID Experiment nr. Species Sample type separation protocol First author Year EV-METRIC
EV200076 1/5 microalgae Cell culture supernatant (d)(U)C Picciotto, Sabrina 2021 67%

Study summary

Full title
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% (92nd 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
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
Cell Name
Dinoflagellate
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.
    • (d)(U)C = (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
Sample Condition
Control condition
EV-producing cells
Dinoflagellate
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Cell number specification
Yes
Separation Method
Differential ultracentrifugation
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
Obtain an EV pellet :
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
Detected EV-associated proteins
beta-actin/ enolase/ HSP70/ Alix
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 Cell culture supernatant (d)(U)C Picciotto, Sabrina 2021 67%

Study summary

Full title
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% (92nd 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
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
Cell Name
Diatom
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.
    • (d)(U)C = (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
Sample Condition
Control condition
EV-producing cells
Diatom
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Cell number specification
Yes
Separation Method
Differential ultracentrifugation
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
Obtain an EV pellet :
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
Detected EV-associated proteins
enolase/ HSP70/ Alix
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 Cell culture supernatant (d)(U)C Picciotto, Sabrina 2021 67%

Study summary

Full title
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% (92nd 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
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
Cell Name
Glaucophyte
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.
    • (d)(U)C = (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
Sample Condition
Control condition
EV-producing cells
Glaucophyte
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Cell number specification
Yes
Separation Method
Differential ultracentrifugation
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
Obtain an EV pellet :
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
Detected EV-associated proteins
Alix/ HSP70/ beta-actin/ enolase
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 Cell culture supernatant (d)(U)C Picciotto, Sabrina 2021 67%

Study summary

Full title
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% (92nd 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
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
Cell Name
Haptophyte
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.
    • (d)(U)C = (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
Sample Condition
Control condition
EV-producing cells
Haptophyte
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Cell number specification
Yes
Separation Method
Differential ultracentrifugation
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
Obtain an EV pellet :
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
Detected EV-associated proteins
Alix/ enolase
Not detected EV-associated proteins
HSP70
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 Cell culture supernatant (d)(U)C Picciotto, Sabrina 2021 67%

Study summary

Full title
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% (92nd 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
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
Cell Name
Chlorophyte
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.
    • (d)(U)C = (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
Sample Condition
Control condition
EV-producing cells
Chlorophyte
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Cell number specification
Yes
Separation Method
Differential ultracentrifugation
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
Obtain an EV pellet :
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
Detected EV-associated proteins
beta-actin/ enolase/ Alix
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
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
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