Synthesis and Identification of Imazalil Antigen

In this study, imazalil (6-[1-(2,4-dichloro-phenyl)-2-(1-imidazolyl) ethoxy] hexanoic acid) was synthesized via substitution reaction between imidazole ethanol and 6-bromo-ethyl followed by hydrolysis reaction to provide a hapten. The synthesized product was identified by thin-layer chromatography (TLC) and 1H-NMR. The hapten was conjugated with bovine serum albumin (BSA) and ovalbumin (OVA) to obtain artificial immune antigen and artificial coating antigen, respectively. The coupling molar ratios were calculated to be 13:1 and 7:1 according to UV spectroscopy analysis, which suggests that artificial antigens are both synthesized successfully. The titer of the antibodies secreted by immunized animals was 1:64000 and the IC50 value was 1.76μ g/L determined by indirect ELISA, which further confirms successful synthesis of artificial antigens. The obtained antibodies can be used for the development of ELISA test kit.