Levels of free anti-(xanthine oxidoreductase) (XOR) antibodies in the serum of normal healthy human subjects were determined, using both human and bovine enzyme as antigen, in an enzyme-linked immunosorbent assay (ELISA). Levels of IgM class anti-(human XOR) antibodies were found to be particularly high (mean values representing approximately 3% of total IgM) and to be significantly higher than levels of IgM anti(bovine XOR) antibodies, indicating that endogenous XOR, rather than ingested bovine milk XOR, is the immunogen. IgM anti-XOR antibody levels were significantly higher in women under 50 years than in age-matched men, or in older women. Levels of IgG class anti-XOR antibodies were much lower and showed no correlation with gender or age. Affinity-purified anti-(human XOR) antibodies only partially inhibited enzymic activities of XOR. The majority of both IgM and IgG anti-(human XOR) antibodies in serum occurred as immune complexes, suggesting that the specific antibodies have a protective role in removing potentially damaging XOR from the circulation.