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Many immunoassays utilize the high affinity interaction of biotin and streptavidin as part of the immunocomplex built to measure the analyte of interest. Therefore, the presence of biotin has the potential to interfere with the assay. Biotin interference on the bead-based Simoa platform was evaluated using Neurofilament-Light (NF-Light™), Neurology 4-Plex A (N4PA: Tau, NF-Light™, GFAP, UCHL1), and Cytokine 3-Plex B (C3PB: IL-6, IL-17A, TNFα).