Enzyme-linked immunosorbent assay for determination of antibodies to xanthine oxidase

An enzyme-linked immunosorbent assay (ELISA) was developed to detect IgG, IgA and IgM antibodies to xanthine oxidase. The method used xanthine oxidase to coat sample wells on microtitre plates. The anti-xanthine oxidase concentrations were determined by reference to standard curves constructed by coating plates with anti-IgG, anti-IgA and anti-IgM to capture antibodies of different classes in standard human serum. The standard curves for IgG, IgA and IgM had a working range of 0 to about 60 ng/ml, and all results with commercial quality control serum fell within expected ranges. The coefficients of variation (CV) for within-batch precision (n = 30) and between-batch precision (n = 20) for IgG and IgM were about 9% and 12% respectively. The detection limit was 2 ng/ml. The ELISA was applied to assay serum samples of 110 Chinese and 110 European healthy subjects. A positively-skewed distribution in their anti-xanthine oxidase IgG and IgM levels was observed.