Development and validation of a radioligand receptor assay for measurement of luteinizing hormone in human serum. 1990

R W Whitcomb, and A L Schneyer
Reproductive Endocrine Unit, Massachusetts General Hospital, Boston 02114.

Studies of circulating LH physiology and pathophysiology are dependent upon measurements of immuno- and bioactivity, both of which have methodologic limitations. We have developed and validated a RRA which allows direct measurement of receptor-bindable LH in human serum. Using a cultured Leydig tumor cell line (MA-10) known to express the CG/LH receptor as the receptor source and polyacrylamide-gel electrophoresis purified hCG as the radioligand, we have established an assay system with the requisite sensitivity (0.04 ng/tube) to measure circulating LH, without significant alteration in total specific binding upon addition of up to 150 microL gonadotropin-free serum when compared to no serum. Standard curves of hLH diluted in gonadotropin-free serum were not statistically different in slope or ED50 from buffer curves. Dilutions of human serum from postmenopausal women and men with Klinefelter's syndrome containing LH measured in the assay were parallel to the standard curve. Further validation of the RRA included measurement of LH by RRA and RIA in daily serum samples from normal women across the menstrual cycle (n = 6) where there was excellent correlation (P less than 0.001) between RRA and RIA measurements with the exception of the mid-cycle surge where the RRA/RIA ratio fell to 0.5. This LH RRA will be useful in further studies of the physiology and biochemistry of LH in human serum.

UI MeSH Term Description Entries
D007986 Luteinizing Hormone A major gonadotropin secreted by the adenohypophysis (PITUITARY GLAND, ANTERIOR). Luteinizing hormone regulates steroid production by the interstitial cells of the TESTIS and the OVARY. The preovulatory LUTEINIZING HORMONE surge in females induces OVULATION, and subsequent LUTEINIZATION of the follicle. LUTEINIZING HORMONE consists of two noncovalently linked subunits, alpha and beta. Within a species, the alpha subunit is common in the three pituitary glycoprotein hormones (TSH, LH and FSH), but the beta subunit is unique and confers its biological specificity. ICSH (Interstitial Cell Stimulating Hormone),Interstitial Cell-Stimulating Hormone,LH (Luteinizing Hormone),Lutropin,Luteoziman,Luteozyman,Hormone, Interstitial Cell-Stimulating,Hormone, Luteinizing,Interstitial Cell Stimulating Hormone
D008183 Luteal Phase The period in the MENSTRUAL CYCLE that follows OVULATION, characterized by the development of CORPUS LUTEUM, increase in PROGESTERONE production by the OVARY and secretion by the glandular epithelium of the ENDOMETRIUM. The luteal phase begins with ovulation and ends with the onset of MENSTRUATION. Menstrual Cycle, Luteal Phase,Menstrual Cycle, Secretory Phase,Menstrual Secretory Phase,Postovulatory Phase,Phase, Luteal,Phase, Postovulatory,Secretory Phase, Menstrual
D008297 Male Males
D008875 Middle Aged An adult aged 45 - 64 years. Middle Age
D011863 Radioimmunoassay Classic quantitative assay for detection of antigen-antibody reactions using a radioactively labeled substance (radioligand) either directly or indirectly to measure the binding of the unlabeled substance to a specific antibody or other receptor system. Non-immunogenic substances (e.g., haptens) can be measured if coupled to larger carrier proteins (e.g., bovine gamma-globulin or human serum albumin) capable of inducing antibody formation. Radioimmunoassays
D011869 Radioligand Assay Quantitative determination of receptor (binding) proteins in body fluids or tissue using radioactively labeled binding reagents (e.g., antibodies, intracellular receptors, plasma binders). Protein-Binding Radioassay,Radioreceptor Assay,Assay, Radioligand,Assay, Radioreceptor,Assays, Radioligand,Assays, Radioreceptor,Protein Binding Radioassay,Protein-Binding Radioassays,Radioassay, Protein-Binding,Radioassays, Protein-Binding,Radioligand Assays,Radioreceptor Assays
D011974 Receptors, LH Those protein complexes or molecular sites on the surfaces and cytoplasm of gonadal cells that bind luteinizing or chorionic gonadotropic hormones and thereby cause the gonadal cells to synthesize and secrete sex steroids. The hormone-receptor complex is internalized from the plasma membrane and initiates steroid synthesis. Chorionic Gonadotropin Receptors,Human Chorionic Gonadotropin Receptors,ICSH Receptors,LH Receptors,LH-hCG Receptor,LH-hCG Receptors,Luteinizing Hormone Receptors,Lutropin Receptor,Lutropin Receptors,Receptors, Chorionic Gonadotropin,Receptors, Human Chorionic Gonadotropin,Receptors, Interstitial Cell-Stimulating Hormone,Receptors, Luteinizing Hormone,hCG Receptors,Chorionic Gonadotropin Receptor,Human Chorionic Gonadotropin Receptor,LH Receptor,Luteinizing Hormone Receptor,Receptors, ICSH,Receptors, Interstitial Cell Stimulating Hormone,Receptors, LH-hCG,Receptors, Lutropin,Receptors, hCG,hCG Receptor,Gonadotropin Receptor, Chorionic,Gonadotropin Receptors, Chorionic,Hormone Receptor, Luteinizing,Hormone Receptors, Luteinizing,LH hCG Receptor,LH hCG Receptors,Receptor, Chorionic Gonadotropin,Receptor, LH,Receptor, LH-hCG,Receptor, Luteinizing Hormone,Receptor, Lutropin,Receptor, hCG,Receptors, LH hCG
D005260 Female Females
D005498 Follicular Phase The period of the MENSTRUAL CYCLE representing follicular growth, increase in ovarian estrogen (ESTROGENS) production, and epithelial proliferation of the ENDOMETRIUM. Follicular phase begins with the onset of MENSTRUATION and ends with OVULATION. Menstrual Cycle, Follicular Phase,Menstrual Cycle, Proliferative Phase,Menstrual Proliferative Phase,Preovulatory Phase,Phase, Follicular,Phase, Menstrual Proliferative,Phase, Preovulatory,Proliferative Phase, Menstrual
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man

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