Abstract:

Two hapaten synthesis methods, succinic anhydride method and alkaline hydrolysis were used for florfenicol deriviazation and the synthesized hapatens, named HP1 and HP2, were separately conjugated with BAS (bovine serum albumin) by active ester method and OVA (ovalbumin) by glutaraldehyde method to obtain two immunoagents, named HP1-BAS and HP2-BAS and two coating antigens, named HP1-OVA and HP2-OVA. HP1-BAS and HP2-BAS were separately used to immunize Balb/c mice for obtaining antisera. Antisera response to HP1-BAS and HP2-BAS were evaluated for their titers using HP1-OVA and HP2-OVA. The mass spectrometric and UV spectral characterization demonstrated that HP1 and HP2 were successfully synthesized. The coupling ratios between HP1 or HP2 and BAS or OVA were determined to be 12.4:1 (HP1-BSA), 8.5:1 (HP1-OVA), 21.8:1 (HP2-BSA) and 15.4:1 (HP2-OVA). The antiserum responses to HP1-BAS had the most specificity to HP2-OVA.

Key words: florfenicol, synthesis of antigen, active ester method, glutaraldehyde method