Synthesis and Identification of Benzoic Acid Artificial Antigen

The ammonolysis method was used to synthesize the artificial antigen of benzoic acid (BA) by phthalic anhydride (PA), the derivative of benzoic acid (BA), in the present study. BA was linked to ovalbumin (OVA) to generate immunogenic antigen BA-OVA and to bovine serum albumin to generate coating antigen BA-BSA, respectively. Artificial antigen was detected and identified by UV and IR spectral studies, as well as SDS-polyacrylamide gels electrophoresis. BA-OVA was further used to immune BALB/c mice to detect the generation of antibody. Ultraviolet spectrophotometry results showed that the ratio of BA to OVA was 174:1. Antibody was detected in serum of immuned mice by ELISA (The titration of the antiserum is 1: 3.2 times 10⁵), which suggested that the antigen induced the specific antibody of BA. These results combined suggested that the artificial antigen of BA was synthesized successfully, which makes it possible to establish monoclonal antibodies.