Characterization of cytokine measurement in human serum, plasma, urine, tears, milk, CSF, and saliva with a MILLIPLEX® multiplex immunoassay

Abstract

Abstract Multiplex immunoassays are valuable research tools for studying circulating cytokines. Here we evaluate the ability of the MILLIPLEX® Human Cytokine/Chemokine/Growth Factor Panel A (Cat. No. HCYTA-60K) 48-plex immunoassay to reproducibly measure cytokines in matched human serum and plasma samples from healthy donors. For comparison, three other bead-based cytokine multiplex kits, featuring many of the same analytes, were analyzed in parallel. Two different lots from each kit were tested with the same matched sample cohort and data were analyzed with Belysa® software. The HCYTA-60K kit was unique in its use of a fixed standard curve, resulting in superior consistency in curve performance between lots. Sample value ratios and correlation demonstrated remarkable consistency of serum and plasma measurement between lots of the HCYTA-60K kit. Further, detectability of additional sample types (urine, tears, milk, CSF, and saliva) was determined with the HCYTA-60K kit. Samples from healthy donors for each biofluid were tested to determine detectability. All analytes could be measured in at least one of the additional sample types. To characterize neuroinflammation during progressive stages of neurological disease, CSF cytokines were measured in samples from mild cognitive impairment, Alzheimer’s Disease, and healthy controls. Our study illustrates the reliability of the MILLIPLEX® HCYTA-60K kit for reproducible measurement of cytokines in serum and plasma between different kit lots. Additionally, we characterize its ability to measure cytokines in urine, tears, milk, CSF, and saliva. These results highlight the utility and robustness of the HCYTA-60K multiplex panel for the quantification of cytokines in diverse biofluids.