Development and Verification of a Novel Multiplex Immunoassay for Immune Profiling in Porcine Models 2257875

Abstract Introduction Multiplex immunoassays play a fundamental role in profiling immune-regulating proteins, including cytokines, chemokines, and growth factors, across both human and animal models. Pigs are particularly valuable in immunology research due to their physiological and genetic similarities to humans, including comparable organ size and immune system function. These similarities make pigs relevant models in translational research. Additionally, their significant presence in the agriculture industry emphasizes the importance of studying porcine health, particularly in the context of livestock management and disease prevention. Methods Here we describe the development of a novel Luminex®-based multiplex immunoassay that can simultaneously detect 32 different cytokines, chemokines, and growth factors in porcine samples. We investigated the expression profiles of these 32 immune mediators across serum and plasma samples from multiple porcine models, including commercial breeds (Yorkshire and German Landrace pigs), and established minipig strains (Hanford, Yucatan, Göttingen, and Sinclair), as well as porcine cell culture supernatants. Results Our findings demonstrate the successful application of this multiplex kit in determining distinct protein expression profiles in serum and plasma samples from multiple porcine breeds and in response to immunostimulants in cell culture models. Conclusion This porcine multiplex immunoassay effectively measures protein expression patterns across diverse porcine breeds and cell lines, supporting both veterinary research and translational studies. Funding Source n/a Topic Categories Veterinary and Comparative Immunology (VET)