A simple, rapid and reliable enzyme-linked immunosorbent assay (ELISA) for measuring estrone-3-glucuronide in urine

A rapid, simple and sensitive enzyme-linked immunosorbent assay for the estimation of estrone-3-glucuronide (E1G) in diluted unprocessed urine has been developed using E1G-penicillinase (E.C.3.5.2.6.) prepared via carbodiimide linkage. Polyvinyl chloride ELISA plate (Dynatech) coated with antibody was used for separation of the antibody bound ligand. A complete assay could be performed within 4 hours with good precision (C.V. less than 17%). Minimum detectable amount of E1G was 25 pg/well. Parallelism was observed between sample dilution curve and standard curve. Recovery of E1G from urine ranged between 85-105%. Results obtained on 29 urine samples by the ELISA procedure were comparable to those obtained with an established RIA procedure (r = 0.96). The present method may thus provide a useful tool for the assessment of ovarian function.