Screening and identification of epitopes of Nfa34810 protein of Nocardia farcinica

ObjectiveTo prepare monoclonal antibodies against Nfa34810 protein of Nocardia farcinica IFM10152 and screen and identify the B cell epitopes.

MethodsFirst, Nfa34810 protein was truncated into P1 and P2 with 20 amino acids overlapping with each other. P1 and P2 were detected to initially map the epitope region by Western Blot using recombinant Nfa34810 protein rabbit polyclonal antibody serum and the mouse polyclonal antibody against N. farcinica IFM10152. Then the P2 where the epitope was located was purified, and BALB/c mice were immunized with the purified P2 and Nfa34810 protein respectively to prepare monoclonal antibodies. The antibodies were identified by Western Blot. Nfa34810 was gradually truncated and expressed and the series of proteins were screened and identified by Western Blot to determine the B cell epitopes of Nfa34810 protein using the prepared monoclonal antibodies by peptide scanning.

ResultsThe monoclonal antibodies of Nfa34810 were prepared and they could react with truncated protein P2 and full-length Nfa34810 protein. One epitope and two epitope regions were successfully screened by peptide scanning.

ConclusionIn this study, the monoclonal antibodies of Nfa34810 protein was successfully prepared and the B cell epitopes were mapped. This study laid a foundation for pathogen detection and epidemiological research of N. farcinica.