Chicken NLRP3 monoclonal antibody development and its antigenic epitope identification

Inflammation is key important pathogenic response that occurs in several poultry diseases. NOD-like receptor protein 3 inflammasome plays a central role in initiating inflammation. However, the lack of antibodies against chicken-derived inflammatory factors presents a major bottleneck in related studies. In this study, we developed a monoclonal antibody (mAb) against chicken NLRP3 using hybridoma technology. This antibody can recognize both eukaryotic and prokaryotic NLRP3 proteins as well as the upregulation of endogenous NLRP3 in HD11 cells induced by lipopolysaccharide or infectious bursal disease virus. Furthermore, a novel antigenic epitope, 42DELEKVTHPSS52, located in the PYD domain of chicken NLRP3 and specifically recognized by this mAb was identified. This epitope is unique to chickens and valuable for distinguishing chicken NLRP3 from orthologs in other species. The developed mAb provides an important tool for detecting chicken NLRP3 and facilitates further study of its antigenic structure and biochemical characteristics.