Plasma progesterone and plasma estradiol levels are commonly used to monitor ovulation in women although for the adequate documentation of ovulation the expense and discomfort of multiple venipuncture sampling may be required. Accurate and definitive information on ovulation in women can be obtained by the simple measurement of metabolites of progesterone and estradiol in early morning urine samples. These analyses have been made possible by the use of our own highly specific monoclonal antibodies to both pregnanediol-3-glucuronide (P-3-G) and estrone-3-glucuronide (E-3-G) and the development of simple, direct, automated enzyme-linked immunosorbent assays. Sequential sampling and the generation of ovulation profiles allows detection of ovulation and identification of the infertile/fertile phases of the cycle for either planned pregnancies or natural family planning. Aberrations in ovulation are easily detected as is documentation of the transition to menopause. The use of sequential, spaced, early morning urine samples for P-3-G and E-3-G allows accurate assessment of ovulatory function rather than relying on the usual single plasma sampling. The data presented also show that the direct determination of plasma pregnanediol-3-glucuronide in plasma is as informative as plasma progesterone measurement.

Adult↗

PubMed1992

Production of a monoclonal antibody to cortisol: application to a direct enzyme-linked immunosorbent assay of plasma.

Cortisol mouse monoclonal antibodies were produced and characterized. Of the four clones studied, supernatant from one clone (A2), compared with other cortisol monoclonal antibodies, showed minimal cross-reactivity to other C21 steroids and was suitable for the direct determination of cortisol in plasma by enzyme-linked immunosorbent assay using a standard 96-well microtiter plate. The enzyme-linked immunosorbent assay uses the immobilized antigen approach, in which cortisol in plasma samples or standards competes with immobilized steroid for antibody-binding sites. After washing, the cortisol antibody bound to the wells of the microtiter plate is detected with antimouse immunoglobulin conjugated to horseradish peroxidase. Following further washing, o-phenylenediamine substrate is added. The enzyme-linked immunosorbent assay is robust and semiautomated. The mean +/- SD recovery from plasma was 97% +/- 6%. Precision studies on three different plasma pools showed mean coefficients of variation of 7.6% and 8.6% for within- and between-assay variation, respectively. The satisfactory performance criteria allow its use in the routine laboratory.

Animals↗