Tonai T, Yokota K, Yano T, Hayashi Y, Yamamoto S, Yamashita K, Miyazaki H
Biochim Biophys Acta. 1985 Oct 2;836(3):335-43. doi: 10.1016/0005-2760(85)90137-7.
A solid-phase enzyme immunoassay of 6-ketoprostaglandin F1 alpha (a stable degradation product of prostaglandin I2) was developed in which the hapten molecule was labeled with beta-galactosidase. The antiserum was bound to a polystyrene tube, and the enzyme-labeled and unlabeled 6-ketoprostaglandin F1 alpha were allowed to react in a competitive manner with the immobilized antibody. The activity of the immunologically bound beta-galactosidase was assayed by fluorometry, and correlated with the amount of unlabeled 6-ketoprostaglandin F1 alpha. According to a calibration curve 6-ketoprostaglandin F1 alpha was detectable in a range of 30 fmol-10 pmol. When 6-ketoprostaglandin F1 alpha was extracted from human serum by using an octadecylsilyl silica column (Sep-Pak C18) and the crude extract was subjected to the enzyme immunoassay, the content of 6-ketoprostaglandin F1 alpha was 50-60 pmol/ml of human serum. An endogenous substance(s) which disturbed the immunoreaction and gave such an apparently high concentration of 6-ketoprostaglandin F1 alpha separated from the endogenous 6-ketoprostaglandin F1 alpha by HPLC. With the purified 6-ketoprostaglandin F1 alpha fraction there was a good correlation (r = 0.994) between enzyme immunoassay and radioimmunoassay. The validity of the enzyme immunoassay was confirmed by gas chromatography-selected ion monitoring.
建立了一种6-酮-前列腺素F1α（前列腺素I2的稳定降解产物）的固相酶免疫测定法，其中半抗原分子用β-半乳糖苷酶标记。抗血清结合到聚苯乙烯管上，酶标记和未标记的6-酮-前列腺素F1α与固定化抗体以竞争方式反应。通过荧光法测定免疫结合的β-半乳糖苷酶的活性，并与未标记的6-酮-前列腺素F1α的量相关。根据校准曲线，6-酮-前列腺素F1α在30 fmol - 10 pmol范围内可检测到。当使用十八烷基硅烷硅胶柱（Sep-Pak C18）从人血清中提取6-酮-前列腺素F1α并将粗提取物进行酶免疫测定时，人血清中6-酮-前列腺素F1α的含量为50 - 60 pmol/ml。一种干扰免疫反应并导致6-酮-前列腺素F1α呈现如此明显高浓度的内源性物质通过高效液相色谱法与内源性6-酮-前列腺素F1α分离。对于纯化的6-酮-前列腺素F1α组分，酶免疫测定法与放射免疫测定法之间具有良好的相关性（r = 0.994）。酶免疫测定法的有效性通过气相色谱 - 选择离子监测得到证实。
