Sensitive enzyme-linked immunosorbent assay for adult T-cell leukemia-derived factor and normal value measurement

Four different monoclonal antibodies against recombinant adult T-cell leukemia-derived factor (ADF), identical to thioredoxin, were established and used for the determination of ADF concentration in serum. Using two of the monoclonal antibodies, we developed a two-step enzyme-linked immunosorbent assay (ELISA) for ADF. This ELISA showed a highly specific reactivity on ADF with no cross-reactivity to several proteins with homologue sequence on the active center. The detection limit of the assay was 2.0 ng/ml (mean +/- 2 SD). The intra- and interassay coefficients of variation (CV) were 0.81-3.74% (n = 8) and 4.78-6.97% (n = 7), respectively. The normal value of ADF mean concentration from 145 healthy donors was 40.8 ng/ml.