Calibration curves obtained for [A] CA125, [B] HE4, [C] MMP-7 and [D] CA72-4 in a singleplex (black circles) and multiplex (white circles) format. Akin to ELISA, the singleplex experiments only employed the antigen of interest and the corresponding antibody for the sandwich immunoassay, whereas the multiplex experiments assayed a cocktail of all four antigens with a cocktail of all four detection antibodies. The multiplex assays for each of the four analytes are superimposable with the individual singleplex assays demonstrating negligible cross-reactivity from the introduction of other reagents due to multiplexing. Representative images (contrast enhanced) of a singleplex assay [E] and a multiplex assay [F], where the singleplex demonstrates specific signal for the analyte of interest (CA125) and the multiplex demonstrates specific signals for a cocktail of analytes.