雌激素调节蛋白pS2竞争性时间分辨免疫荧光分析方法的建立与评估
Development and evaluation of a competitive time-resolved immunofluorometric assay for the estrogen-regulated protein pS2.
作者信息
Rosenberg Zand R S, Jenkins D J, Diamandis E P
机构信息
Department of Nutritional Sciences, Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada.
出版信息
J Clin Lab Anal. 1999;13(5):241-5. doi: 10.1002/(sici)1098-2825(1999)13:5<241::aid-jcla9>3.0.co;2-u.
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 supernatant 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.
我们开发了一种竞争性检测方法来测定雌激素调节蛋白pS2。使用一种单克隆pS2抗体（mAb）和一种生物素化的pS2肽，并采用时间分辨荧光法作为检测技术。该检测方法的检测限为16 ng/mL，且具有精确性（批内和日间变异系数为3 - 12%）。我们使用该检测方法在细胞培养中从时间进程和剂量反应两方面测定了六种甾体激素的活性。当雌二醇作为刺激甾体时，BT - 474乳腺癌细胞系组织培养上清液中的pS2浓度显著更高。观察到雌二醇有显著的时间进程和剂量反应，但其他甾体则没有。一种用于定量pS2的灵敏、可靠且便捷的方法的出现，将使得许多研究应用成为可能，包括筛选天然和合成化合物的潜在雌激素活性。
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