Immunofluorometric assay of prostaglandin D synthase in human tissue extracts and fluids.
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.
Melegos
D N
DN
Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Ontario, Canada.
Diamandis
E P
EP
Oda
H
H
Urade
Y
Y
Hayaishi
O
O
D000328:Adult; D000653:Amniotic Fluid / Q000201:enzymology; D000911:Antibodies, Monoclonal; D001826:Body Fluids / Q000201:enzymology; D001943:Breast Neoplasms / Q000201:enzymology; D005260:Female; D005333:Fetus / Q000201:enzymology; D005348:Fibrocystic Breast Disease / Q000201:enzymology; D015200:Fluoroimmunoassay / Q000379:methods*; D006801:Humans; D019746:Intramolecular Oxidoreductases; D007535:Isomerases / Q000032:analysis*; D054834:Lipocalins; D008297:Male; D008895:Milk, Human / Q000201:enzymology; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011471:Prostatic Neoplasms / Q000201:enzymology; D012661:Semen / Q000201:enzymology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D054834:Lipocalins; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase