Evaluation of an immunoenzymometric assay (IEMA) using automated system for determination of luteinizing hormone and follicle stimulating hormone

It has been proposed that automated systems for immunoenxymometric assay (IEMA) may substitute traditional radioimmunoassay (RIA) for measurement of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in blood due to the advantage of being more rapid, higher sensitivity, lower cost and not requiring radioactive reagents. The study was designed to evaluate both systems using serum samples to determine luteinizing hormone (LH) and follicle stimulatin hormone (FSH) concnetrations. The automatic system (ES-300) for IEMA utilized two monoclonal antibodies, one of them on the solid phase was the specific extractant for the antigen, and the other was a peroxidase labeled antibody which recognizes a different epitope in the antigen molecule, specifically bound in linear proportion to the antigen concentration. Blood samples were obtained from patients who were treated at the hospital for various clinical problems ("problem group") as well as blood samples from patients in whom FSH and LH concentrations were already known ("high", "medium" and "low" levels) by previous RIA ("control group"). IEMA showed a higher sensitivity, 0.42 and 0.96 mIU/ml for FSH and LH, respectively, whereas RIA was 1.95 mIU/ml for both hormones. Intra and interassay coefficient of variation were below 10 percent within the range of 15-50 mIU/ml for FSH and 5-100 mIU for LH; however, the coeffcient of variation was 15 - 25 percent at lower concentrations of FSH and LH. Accuracy of IEMA was evaluated by recovery percentage, thus when high and medium concentrations of FSH and LH were analyzed the recovery was between 99 -104 percent. On the other hand, the recovery was 100 percent when low levels of FSH and LH were used. In coclusion, IEMA resulted reliale when FSH and LH concentrations are in the middle and high range; likewise, the detection limit of IEMA was lower than RIA, particularly for FSH. On the bases of these results, IEMA showed several advantages over RIA, but its reliability diminishes when serum samples contain low FSH and LH concentrations. It is important to extend theses studies to steroid assays and elaborate a database in each laboratory

Monoclonal antibodies specific for the free alpha subunit of glycoprotein hormones and their use in the development of a sensitive immunofluorometric assay

Glicoprotein hormone free alpha subunit has been used as a marker for some pituitary tumors and to study the reactivity of glycoprotein hormone-producing cells under different circunstances. We describe a highly sensitive ans specific immunofluorometric assau for the measurement of serum free alpha subunit levels. The assay is based on a monoclonal antibody, specific for free alpha subunit, bound to microtiter plates. As tracer antibody we employed an europium-labelled free/complexed alpha subunit specific monoclonal antibody. Using overnight incubation and 50µl samples, the least detectable dose was of the order of 4 ng/1. Cross-reactivity with LH, TSH, FSH, and hCG was 6.5, 1.2, 4.3 and 1.1 percent, repectively. Normal adult males showed values ranging from 120 to 790ng/l, not different from normal adult premenopausal females (88 to 604 ng/l). In post-menopausal females, serum concentrations were significantly highler, ranging from 341 to 407 ng/l. In 56 patients with untreated pituitary tumors (18 "non-secreting", 25 GH-producing and 13 prolactin-producing tumors), 10 showed high values, 3 of them from the first group, 3 from the second and 4 from the third. We conclude that this highly sensitive assay can be a valualbe tool for the diagnosis and follow-up of selected patients with pituatary tumors and in other circumstances in which the glycoprotein hormone-producing cells of the pituitary require evaluation

Desarrollo de métodos inmunologicos para regular la fecundidad

A new approach to fertility regulation is the development of vaccines directed against human substances required for reproduction. Potential candidates for immunological interference include reproductive hormones, ovum and sperm antigens, and antigens derived from embryonic or fetal tissue. Several vaccines targeted at the beta chain of the human chorionic gonadotrophin molecule have reached the clinical trial state in Australia, Finland, India and Sweden, and the preliminary results are very encouraging. A prototype vaccine using the beta subunit of ovine luteinizing hormone seems effective in female primates. Contradictory effects on spermatogenisis in male primates have been observed by different researchers using follicle-stimulating hormone as an antigen. Both this and gonadotrophin-releasing hormone require further basic research. Immunological interference could also be aimed at a sperm production or maturation, or at sperm-ovum interaction in the female reproductive tract. The best characterized sperm antigen to date is LDH-C4, an isoenzime of lactic dehydrogenase. The identification of new sperm antigens is being aided by monoclonal antibody techniques. Another approach is to detect, and then mimic, the antisperm antibodies found in women and men with immunologically mediated infertility. Research on ovum antigens is focusing on the zona pellucida (ZP); anti-ZP antibodies