Induction of specific anti-URAT1 antibody after immunization in male C57BL/6J mice. (A) Male diabetic mice were immunized with UIP-1 (n = 10), IA-2(5)-P2-1 (n = 10), or Lipofundin (n = 10), as indicated above. The antibody titer is expressed as the mean OD450 value ± SEM; (B) Antibodies induced by different epitopes in the multi-epitope peptide UIP-1 were identified using relevant coating antigens in ELISA (n = 6/10); (C) Antibody isotypes induced by the different vaccines were determined by ELISA (n = 6/10 for each group); (D) At week 12, sera were collected from UIP-1-immunized mice. Antibodies that specifically recognized and bound to recombinant URAT1 (lane 1) and BSA-conjugated B-cell epitope peptide (lane 4) were detected by Western blotting. BSA (bovine serum albumin) (lane 2) and Asparaginase (lane 3) were not recognized by the antibodies. All data are expressed as the mean ± SEM. * p < 0.05, ** p < 0.01 (UIP-1 vs. placebo group). ##
p < 0.01 (UIP-1 vs. IA-2(5)-P2-1 group).