Enzyme immunoassay of progesterone at the picogram level using beta-galactosidase as label.
We report here the first sensitive enzyme immunoassay of a hapten. A progesterone beta galactosidase conjugate was prepared using carbodiimide as a bifunctional reagent. Rabbit progesterone antisera were previously obtained. The separation of the bound from the free fraction of the label was performed with the help of polymerized anti rabbit gamma-globulins. The enzyme activity of the bound fraction was determined with O-nitrophenyl-beta-D-galactoside as substrate. Specificity and sensitivity (approximately 15 pg) of this enzyme immunoassay can be successfully compared with radioimmunoassay performances. It thus provides a non radioactive, inexpensive and reliable method of small molecule quantitation.
D000818:Animals; D003429:Cross Reactions; D005069:Evaluation Studies as Topic; D005696:Galactosidases; D007118:Immunoassay / Q000379:methods; D007700:Kinetics; D008832:Microchemistry; D011374:Progesterone / Q000032:analysis* / Q000276:immunology; D011485:Protein Binding; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay
D011374:Progesterone; D005696:Galactosidases