Two-hour assay for lutropin during ovulation.

A rapid lutropin assay with a 2-h incubation time and a second antibody/polyethylene glycol separation step is presented. Assay time is shortened by incubating at 37 degrees C and using relatively high concentrations of antibody and radioligand. The interval required for the antigen/antibody reaction varies directly with lutropin concentration, from 1 h for ovulation values to 8 h for low values. After a 2-h incubation, low concentrations have reached 80% of their equilibrium concentration. Separation of the bound fraction by use of combined second antibody/polyethylene glycol (50 g/L) gave one-third the nonspecific binding and as rapid a separation as with polyethylene glycol (180 g/L) alone. Optimal conditions for separating the immune complex were established, and separation was found to be independent of protein concentrations in urine or serum. This tested assay can detect increasing and ovulatory lutropin concentrations in urine or serum, but with some sacrifice in sensitivity.