Luminex assay of plasma IL-6, TNF-α, leptin, and adiponectin: Impact of anticoagulants, single-well analysis, and freeze–thaw stability

BackgroundChronic inflammation from visceral obesity contributes to metabolic disorders and cancer through cytokine imbalance. Accurate cytokine measurement is crucial to understand these links. This study examined challenges in quantifying interleudin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), leptin, and adiponectin in historical biobank samples with limited volumes and varying plasma anticoagulants.MethodsLuminex-MAGPIX platform was used to evaluate three analytical aspects. Single-well precision was assessed via intra- and inter-assay coefficients of variability (CVs). Plasma matrix effects were examined by pairwise comparison of analyte concentrations across citrate, heparin, and EDTA plasma from 10 volunteers. Cytokine stability was tested after one freeze-thaw cycle (analysis of variance).ResultsSingle-well measurements showed good precision (CV