Plasma Biomarkers Measurement Protocols

Plasma CRP was analyzed using a DuoSet enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN, USA). Cytokines were assayed using a premixed Milliplex MAP High Sensitivity Human Cytokine Magnetic Bead Panel (EMD Millipore Corp., St. Charles, MO, USA) and measured using a Luminex 200 instrument (Luminex, Austin, TX, USA). All plasma biomarkers were analyzed by the University of Michigan Cancer Center Immunological Monitoring Core (Ann Arbor, MI, USA). Both methods and detection limits have been described in detail elsewhere [22 (link)]. As with phthalate metabolites, inflammation biomarker measurements below the LOD were replaced by the LOD divided by the square root of 2 [23 ].