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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV210378	9/10	Homo sapiens	MCF-7	(d)(U)C Filtration	Jin D	2018	44%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (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 exosome 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 Filtration Protein markers EV: CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells MCF-7 EV-harvesting Medium Serum free medium 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: speed (g) 110000 Wash: time (min) 70 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Western Blot Antibody details provided? No Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins CD9/ CD63 Detected EV-associated proteins CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 126 EV concentration Yes EM EM-type Transmission-EM Image type Close-up, Wide-field Report size (nm) 90

	EV210378	7/10	Homo sapiens	MCF10A	(d)(U)C Filtration	Jin D	2018	22%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 22% (59th 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 exosome 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 Filtration Protein markers EV: CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells MCF10A EV-harvesting Medium Serum free medium 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: speed (g) 110000 Wash: time (min) 70 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Western Blot Antibody details provided? No Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins CD9/ CD63 Detected EV-associated proteins CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	2/10	Homo sapiens	HepG2	(d)(U)C Filtration	Jin D	2018	14%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 14% (44th 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 exosome 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 Filtration Protein markers EV: CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells HepG2 EV-harvesting Medium Serum free medium 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: speed (g) 110000 Wash: time (min) 70 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Detected EV-associated proteins CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	3/10	Homo sapiens	SGC7901	(d)(U)C Filtration	Jin D	2018	14%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 14% (44th 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 exosome 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 Filtration Protein markers EV: CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells SGC7901 EV-harvesting Medium Serum free medium 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: speed (g) 110000 Wash: time (min) 70 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Detected EV-associated proteins CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	5/10	Homo sapiens	HeLa	(d)(U)C Filtration	Jin D	2018	14%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 14% (44th 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 exosome 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 Filtration Protein markers EV: CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells HeLa EV-harvesting Medium Serum free medium 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: speed (g) 110000 Wash: time (min) 70 Wash: speed (g) 110000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Detected EV-associated proteins CD63/ EpCAM/ CEA/ PTK7/ AFP/ PSMA/ PDGF Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	1/10	Homo sapiens	HepG2	(d)(U)C Filtration	Jin D	2018	0%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 0% (median: 14% 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 exosome 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 Filtration Protein markers EV: EpCAM/ CD63 non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells HepG2 EV-harvesting Medium Serum free medium 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 Pelleting performed Yes Pelleting: speed (g) 20000 Filtration steps > 0.45 µm, Characterization: Protein analysis Protein Concentration Method BCA Detected EV-associated proteins CD63/ EpCAM Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	4/10	Homo sapiens	SGC7901	(d)(U)C Filtration	Jin D	2018	0%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 0% (median: 14% 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 exosome 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 Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells SGC7901 EV-harvesting Medium Serum free medium 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 Pelleting performed Yes Pelleting: speed (g) 20000 Filtration steps > 0.45 µm, Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	6/10	Homo sapiens	HeLa	(d)(U)C Filtration	Jin D	2018	0%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 0% (median: 14% 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 exosome 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 Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells HeLa EV-harvesting Medium Serum free medium 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 Pelleting performed Yes Pelleting: speed (g) 20000 Filtration steps > 0.45 µm, Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	8/10	Homo sapiens	MCF10A	(d)(U)C Filtration	Jin D	2018	0%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 0% (median: 14% 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 exosome 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 Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells MCF10A EV-harvesting Medium Serum free medium 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 Pelleting performed Yes Pelleting: speed (g) 20000 Filtration steps > 0.45 µm, Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis None

	EV210378	10/10	Homo sapiens	MCF-7	(d)(U)C Filtration	Jin D	2018	0%
Study summary Full title ExoAPP: Exosome-Oriented, Aptamer Nanoprobe-Enabled Surface Proteins Profiling and Detection. All authors Jin D, Yang F, Zhang Y, Liu L, Zhou Y, Wang F, Zhang GJ Journal Anal Chem Abstract Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surr (show more...)Tumor exosomes that inherit molecular markers from their parent cells are emerging as cellular "surrogates" in cancer diagnostics. Molecular profiling and detection of exosomes offer a noninvasive access to the state of cancer progression, yet are still technically challenging. Here we report an exosome-oriented, aptamer nanoprobe-based profiling (ExoAPP) assay to phenotype surface proteins and quantify cancerous exosomes in a facile mix-and-detect format. Our ExoAPP interfaces graphene oxide (GO) with target-responsive aptamers to profile exosomal markers across five cell types by complementing with enzyme-assisted exosome recycling, revealing a heterogeneous pattern.This assay achieves a detection limit down to 1.6 × 10 particles/mL, lowered by several orders of magnitude over other homogeneous protocols. Such a sensitive ExoAPP assay allows for monitoring epithelial-mesenchymal transition through heterogeneous exosomes without involving cellular internalization that often occurs in GO-based cargo delivery. Using ExoAPP to analyze blood samples from prostate cancer patients, we find that target exosome can be identified by surface PSMA, suggesting their potential in clinical diagnosis. (hide) EV-METRIC 0% (median: 14% 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 exosome 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 Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim New methodological development Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells MCF-7 EV-harvesting Medium Serum free medium 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 Pelleting performed Yes Pelleting: speed (g) 20000 Filtration steps > 0.45 µm, Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis None

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EV-TRACK ID	EV210378
species	Homo sapiens
sample type	Cell culture
cell type	MCF-7	MCF10A	HepG2	SGC7901	HeLa	HepG2	SGC7901	HeLa	MCF10A	MCF-7
condition	Control condition	Control condition	Control condition	Control condition	Control condition	Control condition	Control condition	Control condition	Control condition	Control condition
separation protocol	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration
Exp. nr.	9	7	2	3	5	1	4	6	8	10
EV-METRIC %	44	22	14	14	14	0	0	0	0	0

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