A nonseparation, time-resolved fluoroimmunoassay to monitor ovarian function and predict potential fertility in women.
The authors describe a nonseparation, time-resolved fluoroimmunoassay involving the use of monoclonal antibodies for the measurement of estrone-3-glucuronide in urine. The method has appropriate sensitivity (12 nmol/l), better specificity than a conventional radioimmunoassay with polyclonal antibodies, and the advantages of speed, simplicity, less imprecision, and improved clinical effectiveness. The labeled antigen is a novel fluorescent europium chelate covalently linked to estrone glucuronide at carbon 6 of the glucuronide moiety. The antibodies were raised against estrone-3-glucuronyl-6-bovine serum albumin. The antibody binding reaction is performed in microtiter wells (or tubes) and involves the addition of labeled antigen in buffer (2 ng/100 microliters), a limited concentration of antibodies in buffer (100 microliters), and standard or urine sample (10 microliters). The mixture is incubated for 15 minutes at room temperature. Time-resolved fluorescence from the unbound label is proportional to the concentration of estrone glucuronide. The method may be used to monitor ovarian function and potential fertility in women.
Barnard
G
G
Department of Obstetrics and Gynaecology, King's College School of Medicine and Dentistry, London, United Kingdom.
O'Reilly
C
C
Dennis
K
K
Collins
W
W
D004970:Estrone / Q000031:analogs & derivatives / Q000652:urine; D005260:Female; D005298:Fertility; D015200:Fluoroimmunoassay / Q000379:methods; D006801:Humans; D010053:Ovary / Q000502:physiology*; D011863:Radioimmunoassay; D012016:Reference Values
C015856:estrone-3-glucuronide; D004970:Estrone