Abstract
Necrotic enteritis (NE) caused by Clostridium perfringens (C. perfringens) has resulted in significant losses for the poultry industry worldwide. Currently, there is no widely promoted vaccine for NE. In this study, immunoprecipitation (IP) was employed to isolate immunogenic proteins of C. perfringens, and 118 potential candidate antigens were identified through liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). From these, three candidate antigen proteins were selected based on their predicted antigenicity, hydrophilicity, stability, and transmembrane signalling properties: ArcB (an ornithine aminotransferase), TmpC (a probable membrane lipoprotein), and EntB (a possible enterotoxin). These three proteins were successfully produced in large quantities using Escherichia coli (E. coli), with confirmed good solubility. Both in vitro and in vivo research demonstrated that these antigens possess strong immunogenicity, eliciting robust antigen-specific humoral and cellular immune responses in chickens and mitigating NE symptoms caused by C. perfringens. The candidate antigens identified through immunoproteomics hold potential as subunit vaccines against C. perfringens infection.