Automated multiplex profiling of human cytokines and autoantibodies using an Automated Assay Workstation

Automated multiplex bead-based immunoassays were performed using an AAW™ Automated Assay Workstation for high-throughput profiling of immune factors and cancer autoantibodies. Human serum cytokine analysis was conducted with MILLIPLEX Human Cytokine Panel A and Panel B kits (48-plex, overnight protocol), and across cytokines spanning a broad dynamic range, automated-to-manual correlation coefficients (R 2 ) were greater than 0.95 with regression slopes between 0.80 and 1.20. Cancer autoantibody profiling used a 15-plex antigen-coated bead multiplex panel on sera from various cancer types and healthy donors (n=96 total samples). The AAW™ workstation automates all critical liquid handling, standard curve preparation, sample transfer, reagent dispensing (matrix solution, Assay Buffer, Standards, QCs, samples, and Beads), and plate setup with robotic lidding. Assay plates were washed off-deck on an automatic plate washer and analyzed with a Luminex instrument. The automated protocol enabled processing of 2-3 plates daily, with streamlined setup which significantly reduce hands-on time. Minimizing manual interventions reduced opportunities for human errors, resulting in greater reliability and efficiency for high-throughput applications. The AAW™ performance confirms robust, reproducible, and scalable multiplex analysis for cancer and immunology research.