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Fig. 4 | Biomarker Research

Fig. 4

From: Selective isolation of extracellular vesicles from minimally processed human plasma as a translational strategy for liquid biopsies

Fig. 4

Direct EV subset detection and quantification on beads using fluorescently-tagged antibodies. (A) Increasing numbers of HT29 EVs captured from PBS-BSA with triple-coated beads were detected by staining IP complexes with CD9-PE. PE fluorescence signals highly correlated with the number of spiked and recovered EVs, evidenced by linear regression analysis (R2 = 0,9992). (B) Scalar amounts of HT29 EVs were spiked in “EV-depleted plasma” (donor 7) and recovered using triple-coated beads, followed by staining with CD9-PE. A linear trendline showed correlation between the amount of spike and S/N obtained with CD9-PE, from 1 × 108 to 1 × 107 EVs (R2 = 0,9783). An outlier mean S/N value at 5 × 106 spiked EVs was presented in red. (C) Platelet-derived EVs were isolated from plasma samples of 3 different donors using anti-CD61 beads. Target subpopulations (CD41-PE) were detected against a negative control antibody (IgG1k-PE). (D) Cryo-TEM image of platelet-derived EVs recovered from plasma with anti-CD61 beads. Scale bars are shown on the top right corner of the image

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