Improvement of estradiol enzymoimmunoassay, using a monoclonal antibody and an avidin/biotin amplification system.
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.
de Lauzon
S
S
U.E.R. Biomédicale des Saints-Pères, Paris, France.
el Jabri
J
J
Desfosses
B
B
Cittanova
N
N
D000911:Antibodies, Monoclonal; D001360:Avidin; D001710:Biotin; D004958:Estradiol / Q000032:analysis*; D005069:Evaluation Studies as Topic; D007124:Immunoenzyme Techniques
D000911:Antibodies, Monoclonal; D001360:Avidin; D004958:Estradiol; D001710:Biotin