Magnetic bead-based 28-plex immunoassay for the quantification of rhesus and cynomolgus monkey cytokines (144.34)

Abstract

Non-human primates possess high degrees of homology with humans, and thus are crucial model systems in a number of areas of biomedical research (neuroscience, endocrinology, reproductive physiology, cardiovascular physiology, infectious diseases, vaccine and drug efficacy and safety). Monitoring complex cytokine profiles has proven to be invaluable in these various areas. Using the Luminex® xMAP® platform, Life Technologies has developed a 28-plex magnetic bead-based cytokine immunoassay for the analysis of rhesus monkey (Macaca mulatta) and cynomolgus monkey (Macaca fascicularis) samples. Validated sample types include serum, plasma, and conditioned tissue culture medium. Analytes include: EGF, eotaxin, FGF basic, G-CSF, GM-CSF, HGF, IFN-γ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 p40/p70, IL-15, IL-17, I-TAC, MDC, MIF, MIG, MIP-1α, MIP-1β, MCP-1, RANTES, TNF-α, and VEGF. This magnetic-bead based assay is compatible with robotic washing systems, and thus may be fully automated. Total incubation time is 3.5 hours. The utility of the assay is demonstrated by determining cytokine profiles obtained with LPS-stimulated or PMA plus A23187-stimulated peripheral blood mononuclear cells (PBMCs) from rhesus monkeys and cynomolgus monkeys. Linearity of dilution experiments with tissue culture medium and serum produce R2 values of 0.98 and greater, while sample recovery routinely falls between 80 and 120%. Intra-assay and inter-assay precision is ≤10% CV.