Abstract
To determine whether multiplex immunoassays yield results that are comparable to those obtained with traditional enzyme-linked immunosorbent assays (ELISAs), cytokine concentrations were determined in 53 EDTA plasma samples from 7 college-age healthy individuals (3 female) undergoing experimental placebo ( n = 4) or endotoxin ( n = 3) exposure; 6–8 blood samples per subject were collected over a 7-h observation period. Plasma was assayed according to the manufacturer’s protocols for IL1-β, IL6, and TNF-α by high sensitivity ELISAs (Quantikine, R& D Systems), and as part of an 11-plex Performance High Sensitivity Human Cytokine multiplex bead-based (Luminex) assay (R& D Systems). When all samples in the placebo and endotoxin conditions were examined together, strong correlations were seen between ELISA and multiplex concentrations of IL1-β ( r = 0.91), IL6 ( r = 0.99), and TNF-α ( r = 0.94; all p values r = 0.89) and IL6 ( r = 0.98; both p values r = −0.28). This difference for TNF was not a function of assay sensitivity, as the calculated TNF concentrations for all placebo samples were within the range of the standard curve on both ELISA and multiplex assays. For IL-1 and IL-6, ELISAs and multiplex immunoassays yield similar results for samples in the physiological circulating range.
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