Development of NULISAseq™ Inflammation Panel 250 for Comprehensive Immune Profiling

ABSTRACT
Comprehensive profiling of cytokines and chemokines in blood can provide deeper insights into the mechanisms underlying highly complex and heterogeneous groups of autoimmune diseases. However, many of these proteins are present at very low concentrations in plasma, below the limit of detection of current immunoassays. We recently developed an automated multiplex immunoassay technology, NULISA, capable of attomolar-level sensitivity and high levels of multiplexing, and we demonstrated the performance characteristics of a 200-plex assay and its correlation with existing platforms2. Here we report the development of an expanded inflammation-focused panel called NULISAseq™ Inflammation Panel 250 (IP250) targeting 200 cytokines/chemokines and their receptors and 50 other important inflammation and immune response-related proteins. The assay demonstrated high sensitivity, detecting 97.6% of the targets in at least 50% of plasma samples, and high precision, with median intra-plate CV of <15% and inter-plate CV of <15%. It also demonstrated the ability to quantify proteins of a wide dynamic range without diluting samples. Comprehensive testing for target specificity demonstrated that 95% of the assays in the panel had <1% cross reactivity to nonspecific targets. In summary, with ultrahigh sensitivity and the most comprehensive coverage of inflammatory cytokines/chemokines, the Inflammation Panel 250 assay provides a powerful discovery tool for inflammation and immunology research at the systems level, which may lead to new diagnostic biomarkers and therapeutic targets.