Figure 4

Characterization of exosomes isolated using PEG/SEC methodology from porcine sera. Bradford and flow cytometry analyses (A), Sandwich ELISA for exosomes derived from non-viremic animal (B), are represented. MFI: Median fluorescence unit, mg/mL: Bradford measure unit, CD63-CD81 are molecular markers for exosomes characterization. For FACS and Bradford analyses is evident that higher values are represented in comparison with non-concentrated samples (10-fold change). In addition, the indirect quantification through Bradford of protein associated with exosomes it is possible when sera is concentrated through PEG and separated using SEC, making this an important tool for further evaluation of immunogenic properties in vitro and in vivo. For the sandwich ELISA, exosomes derived from non-infected animal were tested against two types of porcine sera. Optical density (OD) was measured at 450 nm and represented in the “Y” axis. “X” axis shows the dilution factor for exosomes samples (1 to 1/200). Duplicated values are represented as squares and dots and mean as a line.