Two-hour Sephadex column method for assay of unconjugated estriol in serum

The column method of Christner and Fetter (Steroids 24: 327, 1974) has been modified to give a simple, rapid assay for unconjugated estriol in serum. Estriol is isolated from serum by being retained on a Sephadex column, while estriol conjugates and serum proteins are eluted. The sample and labeled estriol compete for antibody on the column. Antibody is eluted, removing proportional amounts of sample and labeled estriol. Our modifications include using stable serum-based standards, shortening the incubation to 10 min, complete removal of conjugates, and regeneration of the columns so that they can be used repeatedly. The assay, which can be completed in 2 h, has a mid-range interassay CV of 8.2%.