Abstract: Aiming at the problem that the small molecule uric acid lacked immunogenicity and it was difficult to directly detect it using immunological methods, we modified uric acid with succinic anhydride to obtain a modified uric acid hapten, and then conjugated the uric acid hapten with bovine serum albumin and ovalbumin to prepare uric acid artificial antigens and coating antigens. We immunized mice with uric acid artificial antigens to prepare uric acid polyclonal antibodies. Uric acid hapten was characterized by Fourier transform infrared spectroscopy and mass spectrometry, artificial antigen and coating antigen were characterized by ultraviolet spectrophotometry and polyacrylamide gel electrophoresis, and antibody titer, sensitivity and specificity were determined by indirect competitive enzyme-linked immunosorbent assay (ELISA). The results show that the titers of the multi antibody serum are all above 1∶25 600, and the sensitivities of the antibodies in mice 1—4 are 10.337,8.426,16.457,21.177 μg/mL, respectively. The cross reactivity rates between the antibodies and structural analogues of uric acid are all less than 1%. It can be seen that the uric acid polyclonal antibody prepared by immunization in this experiment has achieved high potency, high sensitivity, and strong specificity, providing experimental basis for the immunological detection of uric acid. 

Key words: uric acid, hapten, artificial antigen, polyclonal antibody