Multiplex Immunoassay Profiling of Blood Samples

The ProcartaPlex multiplex immunoassay kit was purchased from ThermoFisher Scientific and U-plex array was purchased from Meso Scale Diagnostics (MSD). Blood samples were collected at the end of the experiment from the control and experimental groups. Plasma or serum samples were collected at the time of sacrifice and separated from whole blood following the manufacturer’s instructions. Briefly, for the ProcartaPlex assay, antigen standards were prepared and incubated with magnetic beads. Plasma samples were incubated with the beads for 1–2 h. After washes with the wash buffer, samples were incubated with detection antibody mixture for 30 min at room temperature with shaking. Streptavidin labeled with PE fluorophore was later added to the samples followed by incubation for 30 min. The plate was analyzed further on a Luminex instrument. Samples were subjected to a U-plex array as described^{8 (link)}.