Abstract:To study the preparation of polyclonal antibody against Ebp fimbriae subunit protein of animal-originated Enterococcus faecalis and its possible immunological effect.The antigenic epitopes of EbpA, EbpB and EbpC proteins of E. faecalis fimbriae were predicted by bioinformatics software. Six antigenic epitopes were found, and the recombinant plasmid was constructed, expressed and purified. After immunizing New Zealand white rabbits, six fimbriae subunit polyclonal antibodies were obtained, and then the antibody specificity and biofilm blocking effect were analyzed.The six fimbriae subunit recombinant proteins were consistent with the predicted molecular weight of the target proteins. All of them had immunogenicity to New Zealand white rabbits. The titers of polyclonal antibodies against EbpA1、EbpA2、EbpA3、EbpB1、EbpC1and EbpC2 subunit proteins were 1∶16, 1∶8, 1∶32, 1∶64 and 1∶64 by bi-directional immune-agar diffusion test respectively. WB detection and biofilm blocking test showed that the six polyclonal antibodies had antigen specificity and biofilm blocking effect of wild strains, and the biofilm blocking effect of EbpA1, EbpC1 and EbpA3 was the strongest. In conclusion, EbpA1, EbpC1 and EbpA3 subunit proteins of Enterococcus faecalis may lay the foundation for Enterococcus faecalis immune-prevention and treatment.
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