Construction, expression and purification of a novel CadF–based multiepitope antigen and its immunogenic polyclonal antibody specific to Campylobacter jejuni and Campylobacter coli

Abstract

Campylobacteriosis is a disease in humans caused by the infection from Campylobacter spp. Human cases are mainly due to Campylobacter jejuni, although C. coli can cause gastroenteritis in humans as well. The bacteria are commensal in chicken tract and can be contaminated into chicken products during processing. Obviously, detecting reagents such as a specific antibody is essential for the development of immune–based detection methods for C. jejuni or C. coli. In this study, in silico techniques were used to design a chimeric recombinant antigen, named multiepitope antigen (MEA), for the production of specific polyclonal antibody. To design MEA polypeptide based on C. jejuni fibronectin–binding protein or CadF, four conserved and unique antigenic peptides were identified and fused together directly. The C. jejuni CadF–based MEA polypeptide fused with two single six–histidine tags at both C– and N–terminal ends was expressed under Escherichia coli expression system. The recombinant MEA was successfully produced and purified by Ni–NTA resin with a high satisfactory yield. Indirect ELISA results showed that anti–MEA polyclonal antibody derived from rabbit serum had a titer of 16,000, indicating high antigenicity of MEA polypeptide. Dot blot results also confirmed that the produced anti–MEA antibody could specifically recognize both C. jejuni and C. coli whole cells as expected while there was no cross–reactivity to non–Campylobacter spp. tested in this study.