Milliplex map for luminex technology

Serum CC16 levels were measured in samples from both early school-age and adolescence at the Asthma and Airway Disease Research Center Immunology Lab at the University of Arizona using a commercially available ELISA kit (BioVendor, Asheville, NC and Modrice, Czech Republic).
Serum levels of four inflammatory biomarkers previously associated with CF lung disease severity(25 (link)) were measured in samples from adolescence in the Pediatric Clinical Translational Research Center Core Laboratory at Children’s Hospital Colorado and University of Colorado Anschutz Medical Campus (Aurora, CO), which serves as the Center for Biochemical Markers for the CF Foundation. Proteins were measured using validated commercially available assays: high sensitivity C reactive protein (hsCRP; Siemens Nephelometer assay, Siemens Healthcare, Tarrytown, NY), serum amyloid A (SAA; Milliplex MAP® for Luminex Technology, EMD Millipore, St. Charles, MO), calprotectin (ALPCO Diagnostics, Salem, NH), and granulocyte-colony stimulating factor (G-CSF; Milliplex MAP® for Luminex Technology, EMD Millipore, St. Charles, MO). The lower limits of detection for these assays were: hsCRP, 0.007 mg/L; SAA, 500 ng/mL; calprotectin, 0.4 μg/mL, and G-CSF, 14 pg/mL. All assays were performed in duplicate and mean values were used for analysis.

Serum aliquots were analyzed using validated commercially available assays for the following protein biomarkers: high sensitivity C reactive protein (hsCRP; Siemens Nephelometer assay, Siemens Healthcare, Tarrytown, NY), serum amyloid A (SAA; Milliplex MAP® for Luminex Technology, EMD Millipore, St. Charles, MO), calprotectin (American Laboratory Products Company-ALPCO, Salem, NH), and myeloperoxidase (MPO; ELISA, R&D systems, Minneapolis, MN). The lower limits of detection for these assays were: hsCRP, 0.007 mg/L; SAA, 500 ng/mL; calprotectin, 0.4 μg/mL; and MPO, 15 ng/mL. The intra-assay coefficients of variation (CV) were less than 15% for all assays. The sputum supernatants were analyzed for the following markers of inflammation: free neutrophil elastase activity (NE) (spectrophotometric assay based on the hydrolysis of the specific substrate MeO-suc-Ala-Ala-Pro-Ala-p-nitroanilide [Sigma Chemical Co, St. Louis, MO]), MPO (ELISA, R&D systems, Minneapolis, MN), IL-8 and TNF-α (Luminex FluorokineTechnology, R&D Systems, Minneapolis, MN). The lower limits of detection for these assays were: free neutrophil elastase activity, 8 μg/mL; MPO, 120 ng/mL; IL-8, 3, 120 pg/mL; and TNF-α, 12.5 pg/mL. All assays were performed in duplicate and mean values were used for analysis.