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Improvement of estradiol enzymoimmunoassay, using a monoclonal antibody and an avidin/biotin amplification system
S de Lauzon1, J el Jabri, B Desfosses
1U.E.R. Biomédicale des Saints-Pères, Paris, France.
Abstract:
A microtitre plate enzyme immunoassay for estradiol, using a purified monoclonal antibody covalently bound to peroxidase and a small amount of immobilized immunogen, was optimized. Decreasing the antibody concentration to 2 X 10(-10) M (Kd/5) gave optimum estradiol detectability. The enzymatic signal was, however, very low in this assay. A 14-fold enhancement could be obtained using an avidin-biotin system in which several biotin molecules are conjugated with the antibody, providing multiple sites for binding by an avidin-enzyme complex. Further reagent concentration optimization gave an assay in which a range of 2 to 140 pg estradiol/well could be assayed simply and reproducibly.