Barnard G, O'Reilly C, Dennis K, Collins W
Department of Obstetrics and Gynaecology, King's College School of Medicine and Dentistry, London, United Kingdom.
Fertil Steril. 1989 Jul;52(1):60-5. doi: 10.1016/s0015-0282(16)60789-6.
The authors describe a nonseparation, time-resolved fluoroimmunoassay involving the use of monoclonal antibodies for the measurement of estrone-3-glucuronide in urine. The method has appropriate sensitivity (12 nmol/l), better specificity than a conventional radioimmunoassay with polyclonal antibodies, and the advantages of speed, simplicity, less imprecision, and improved clinical effectiveness. The labeled antigen is a novel fluorescent europium chelate covalently linked to estrone glucuronide at carbon 6 of the glucuronide moiety. The antibodies were raised against estrone-3-glucuronyl-6-bovine serum albumin. The antibody binding reaction is performed in microtiter wells (or tubes) and involves the addition of labeled antigen in buffer (2 ng/100 microliters), a limited concentration of antibodies in buffer (100 microliters), and standard or urine sample (10 microliters). The mixture is incubated for 15 minutes at room temperature. Time-resolved fluorescence from the unbound label is proportional to the concentration of estrone glucuronide. The method may be used to monitor ovarian function and potential fertility in women.
作者描述了一种非分离的时间分辨荧光免疫分析方法，该方法使用单克隆抗体来测量尿液中的雌酮-3-葡萄糖醛酸苷。该方法具有适当的灵敏度（12纳摩尔/升），比传统的多克隆抗体放射免疫分析具有更好的特异性，并且具有速度快、操作简单、精密度低和临床效果改善等优点。标记抗原是一种新型的荧光铕螯合物，在葡萄糖醛酸部分的碳6处与雌酮葡萄糖醛酸共价连接。这些抗体是针对雌酮-3-葡萄糖醛酰-6-牛血清白蛋白产生的。抗体结合反应在微量滴定孔（或试管）中进行，包括在缓冲液中加入标记抗原（2纳克/100微升）、缓冲液中有限浓度的抗体（100微升）以及标准品或尿液样本（10微升）。混合物在室温下孵育15分钟。未结合标记物的时间分辨荧光与雌酮葡萄糖醛酸苷的浓度成正比。该方法可用于监测女性的卵巢功能和潜在生育能力。
