Taylor and Francis Group, Journal of Immunoassay and Immunochemistry, 3(24), p. 273-283
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Cytokines are often measured using ELISAs and chemiluminescence (CMIL) is reported to exhibit increased sensitivity compared to colorimetric (COL) assays. CMIL also has a wider dynamic detection range. We sought to directly compare ELISAs for measuring human TNF and IL-8 using CMIL or COL. CMIL substrates with glow fluorescence were obtained from 4 different commercial sources while the COL substrate was TMB. ELISAs for TNF and IL-8 were run under identical conditions and the standard curve extended from 0.5 to 4000 pg/ML. The COL substrate demonstrated a sigmoid shaped curve when plotted on a log-linear scale while the CMIL continued to increase up to the highest concentration. Both substrates were modeled most accurately by a 4 parameter equation with R values > 0.99. The standard curves for both the IL-8 and TNF demonstrated a lower limit of detection (LLD) for the COL comparable to the CMIL detection system. To precisely define the LLD quadruplicate blanks were run and the mean plus 4 standard deviations were used. By these criteria, the COL assay routinely had a LLD of < 1.5 pg/ML which was better than any of the CMIL substrates. Our data demonstrate the COL assays have the same or better sensitivity than CMIL and are significantly less expensive.