Melegos D N, Diamandis E P, Oda H, Urade Y, Hayaishi O
Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Ontario, Canada.
Clin Chem. 1996 Dec;42(12):1984-91.
A two-site sandwich-type assay for human prostaglandin D (PGD) synthase (beta-trace) was developed with two monoclonal antibodies and using time-resolved fluorometry as the detection technique. The assay is precise (CVs < 10%), accurate, and highly specific for PGD synthase and has a detection limit of 0.05 microgram/L. Using this assay, we measured PGD synthase concentrations in serum, urine, amniotic fluid, cerebrospinal fluid (CSF), seminal plasma, breast cyst fluid, breast discharge fluid, breast milk, and breast tumor extracts. The highest concentrations were found in CSF. We identified proteolytic degradation of PGD synthase in amniotic fluid. Fetal tissues contained various amounts of the enzyme, with the highest values being found in brain and heart. In placental extracts, PGD synthase content was greatest at 11-28 weeks of gestation-in accordance with the concentrations measured in amniotic fluids for this gestational period. We conclude that PGD synthase is ubiquitous and is present in many fluids and tissues of adults and fetuses. This first quantitative and sensitive assay of PGD synthase should facilitate expansion of knowledge on this enzyme and possibly will have applications for diagnosis and monitoring of human diseases.
采用两种单克隆抗体，以时间分辨荧光法作为检测技术，开发了一种用于检测人前列腺素D（PGD）合酶（β-微量蛋白）的双位点夹心型检测方法。该检测方法精确（变异系数<10%）、准确，对PGD合酶具有高度特异性，检测限为0.05微克/升。利用该检测方法，我们测定了血清、尿液、羊水、脑脊液（CSF）、精浆、乳腺囊肿液、乳腺分泌液、母乳和乳腺肿瘤提取物中的PGD合酶浓度。脑脊液中的浓度最高。我们发现羊水中PGD合酶存在蛋白水解降解。胎儿组织含有不同量的该酶，在脑和心脏中含量最高。在胎盘提取物中，PGD合酶含量在妊娠11 - 28周时最高，这与该妊娠期羊水中测得的浓度一致。我们得出结论，PGD合酶广泛存在于成人和胎儿的许多体液和组织中。这种首次对PGD合酶进行的定量且灵敏的检测方法应有助于扩展对该酶的认识，并且可能在人类疾病的诊断和监测中得到应用。
