Preparation of monoclonal antibodies against pyruvate dehydrogenase E1 component beta subunit (PDHB) protein of Mycoplasma bovis and the identification of its two novel B-cell epitopes.

Mycoplasma bovis (M. bovis) is a significant pathogen causing mastitis, pneumonia, and other diseases in dairy cattle. This pathogen severely affects the health and productivity of dairy cattle, thereby resulting in substantial economic losses for the dairy industry. Developing highly specific and sensitive diagnostic reagents, alongside safe and effective vaccines, is a critical strategy for controlling and preventing M. bovis infections. The pyruvate dehydrogenase β subunit (PDHB) protein, known for its high conservation and strong immunogenicity, has been identified as a promising target for diagnostics and vaccine development. In this study, we generated two monoclonal antibodies against the PDHB protein, designated B15-41 and E20-5, and precisely identified their respective B-cell epitopes for the first time: amino acid 10GALNHA15 and 52DQRVWD57. Further bioinformatics analysis revealed that these epitopes are highly conserved among selected reference strains and are exposed on the PDHB protein surface, facilitating recognition by the immune system. These findings contribute to our further understanding of the structural and functional characteristics of the PDHB protein and advance the development of specific detection methods against M. bovis.