Figure 5.

Multiplex bead-based flow cytometry assay: antigen profiling of EVs from PFP samples from leukemia patients at diagnosis and HBDs: (A) Experimental outline of multiplex bead-based detection of EV antigen profiles. The concentration of PFP-derived EVs was determined prior to use in the assay by either total protein count or NTA and defined doses of 50 µg or 5 × 10⁸ EVs, respectively, were used as assay inputs. (B) Representative dot plots showing the negative control (MBP buffer), as well as a sample from a HBD, an AML and a B-ALL patient. (C) Median APC fluorescence intensities (MFI) for all 39 bead populations after background correction (MBP buffer values subtracted from measured PFP values). Mean ± SEM of the three independent measurements (replicates) are shown (n = 1). For improved comparability of plots with different axis scaling, we included an arbitrary dotted line at an MFI value of 10.