Development and evaluation of a competitive time-resolved immunofluorometric assay for the estrogen-regulated protein pS2

✍ Scribed by Rachel S. Rosenberg Zand; David J.A. Jenkins; Eleftherios P. Diamandis

• Book ID: 101270817
• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 73 KB
• Volume: 13
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/(sici)1098-2825(1999)13:5<241::aid-jcla9>3.0.co;2-u

✦ Synopsis

We have developed a competitive assay to measure the estrogen-regulated protein pS2. A monoclonal pS2 antibody (mAb) and a biotinylated pS2 peptide are used, with time-resolved fluorometry as a detection technique. The assay has a detection limit of 16 ng/mL and is precise (within-run and day-to-day CVs 3-12%). We used this assay to determine steroid hormone activity of six steroids in cell culture, both in terms of time course and dose response. pS2 concentrations in the tissue culture superna-tant of the BT-474 breast carcinoma cell line were significantly higher when estradiol was the stimulating steroid. There was a significant time course and dose response observed for estradiol, but not for the other steroids. The availability of a sensitive, reliable, and convenient method for quantifying pS2 will allow for many research applications including the screening of natural and synthetic compounds for putative estrogenic activity.