Figure 1
Particle and protein yields of uEVs isolated by different methods. uEVs were isolated from 50 ml of the same urine by 5 different methods: UC = ultracentrifugation, UC-SEC = ultracentrifugation followed by size exclusion chromatography, C-SEC = concentration followed by size exclusion chromatography, PEG = isolation by polyethylene glycol, PEG-SEC = isolation by polyethylene glycol followed by size exclusion chromatography. The results are shown as mean ± SEM of 5 experiments performed in triplicates. (A) Number of uEVs isolated by 5 different methods. NTA was performed on isolated uEVs to calculate total particles (left Y-axis in red) and particles in range of 50–150 nm (right Y-axis in blue) in the same sample. (B) Size characteristics of uEVs. Mean, mode, standard deviation and different size distributions (D10, D50 and D90) were calculated in samples isolated by 5 different methods. D10 = 10% of particles are below the size indicated as D10, D50 = 50% of particles are below the size indicated as D50, D90 = 90% of particles are below the size indicated as D90. Tukey’s multiple comparisons test showed the most significant differences in D90 between PEG-SEC and all other methods (adjusted p value < 0.1–0.0001). (C) Protein content of uEVs measured by BCA. Tukey’s multiple comparisons test was performed and adjusted p-value of UC vs UC-SEC was 0.119, for UC vs C-SEC was 0.0707 and for UC vs. PEG-SEC was 0.0150. (D) Correlation between the total protein amount (left Y-axis in red) and 50–150 nm particle count (right Y-axis in blue) of uEVs.
