Lewis J G, Clifford J K, Elder P A
Department of Clinical Biochemistry, Christchurch Hospital, New Zealand.
Steroids. 1990 Jul;55(7):314-8. doi: 10.1016/0039-128x(90)90035-a.
Monoclonal antibodies to pregnanediol-3-glucuronide were produced and characterized. One of three clones investigated provided antibody suitable for a direct urinary enzyme-linked immunosorbent assay (ELISA). The ELISA uses a pregnanediol-thyroglobulin conjugate adsorbed onto the wells of a standard 96-well microtiter plate. Pregnanediol-3-glucuronide in standards or diluted urine competes with the immobilized steroid for antibody-binding sites. After washing, mouse monoclonal antibody bound to the plate is probed with antimouse immunoglobulin peroxidase. After further washing, o-phenylenediamine substrate is added and, finally, the absorbance is read at 492 nm. The ELISA shows excellent performance and agreement with the previous gas chromatographic method. The ELISA is ideal for aiding the assessment of ovarian function in the routine laboratory.
制备并鉴定了针对孕二醇 - 3 - 葡萄糖醛酸苷的单克隆抗体。在所研究的三个克隆中，有一个克隆产生的抗体适用于直接尿酶联免疫吸附测定（ELISA）。该ELISA使用吸附在标准96孔微量滴定板孔上的孕二醇 - 甲状腺球蛋白偶联物。标准品或稀释尿液中的孕二醇 - 3 - 葡萄糖醛酸苷与固定化类固醇竞争抗体结合位点。洗涤后，用抗小鼠免疫球蛋白过氧化物酶检测与板结合的小鼠单克隆抗体。进一步洗涤后，加入邻苯二胺底物，最后在492nm处读取吸光度。该ELISA表现出优异的性能，与先前的气相色谱法结果一致。该ELISA非常适合在常规实验室中辅助评估卵巢功能。
