Fig. 2

Overview of methods for cytokine profiling using Luminex MILLIPLEX® assay. Plasma contains an unspecified concentration of analytes of interest, i.e., cytokines, chemokines, interleukins, and growth factors. To determine the concentrations of the analytes, each Luminex well contains multiple types of color-coded beads conjugated to a capture antibody that binds the specific analyte of interest. In presence of the analyte, a biotin-conjugated detection antibody will bind to the capture antibody-analyte complex. Adding phycoerythrin-labeled streptavidin will produce fluorescence that is color-specific for each analyte and allows for measuring the mean fluorescence intensity (MFI) of each analyte in each Luminex well. Comparing MFI at different dilution levels with standard solutions allows for determining the concentrations of the analytes of interest