Color-coded beads, pre-coated with analyte-specific capture antibody for the molecule of interest, are added. Multiple analytes can be simultaneously detected in the same sample.

Analyte-specific antibodies capture the analyte of interest. Biotinylated detection antibodies specific to the analyte of interest are added and form an antibody-antigen sandwich.

Phycoerythrin (PE)-conjugated Streptavidin is added.

The beads are read a dual-laser flow-based detection instrument, such as the Luminex 200™ or Bio-Rad® Bio-Plex® analyzer. One laser classifies the bead and determines the analyte that is being detected. The second laser determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of bound analyte.