Abstract: To develop an immunoassay for melamine residues, melamine hapten MEA and MEB were synthesized by the reaction between 2-Chloro-4,6-diamino-1,3,5-triazine and aminocaproic acid or p-aminobenzoic acid, respectively. Then two artificial antigens (MEA-BSA and MEB-OVA) were prepared by active ester method (AE) and mixed anhydride method (MA) respectively. Mass spectral analysis demonstrated the successful synthesis of MEA and MEB. Characterization of MEA-BSA and MEB-OVA was conducted by using UV spectrophotometric method. The numbers of hapten molecules per protein molecule estimated by ultraviolet spectrophotometry according to the absorbance at 253.5 nm for two conjugates were 25.28 and 13.11 respectively. The results suggested that the artificial antigens can be used for preparing specificity anti-melamine antibody.

Key words: melamine, hapten, artificial antigen