DEXTRAN-COATED CHARCOAL USED IN THE RADIOIMMUNOASSAY OF HUMAN PITUITARY LUTEINIZING HORMONE

1973 ◽  
Vol 73 (3) ◽  
pp. 444-454 ◽  
Author(s):  
T. Sand ◽  
P. A. Torjesen
Keyword(s):  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Protein Composition ◽  
Thyroid Stimulating Hormone ◽  
Human Chorionic Gonadotrophin ◽  
Human Thyroid ◽  
Serum Samples ◽  
Human Pituitary ◽  
The Difference ◽  
Stimulating Hormone

ABSTRACT A radioimmunoassay for human pituitary luteinizing hormone (LH) using charcoal for the separation of free from antibody-bound hormone is described. The ability of the various types of charcoal preparations tested to separate free from antibody-bound hormone differed greatly as did the amount required to give maximum adsorption of free hormone. It was also found that the adsorption of free and antibody-bound hormone was greatly influenced by the presence of other proteins. Hence it was necessary to add human serum to the standard tubes before the addition of the charcoal-dextran suspension, in order to compensate for the difference in protein composition between the standards and the serum samples. Two antisera obtained from rabbits immunized with human chorionic gonadotrophin (HCG) were used. One of the antisera had an affinity to human thyroid-stimulating hormone (TSH) and human follicle-stimulating hormone (FSH) of less than 10% as compared to that of LH, while the other had an affinity of about 30 % as compared to that of LH.

PREPARATION AND PROPERTIES OF SUBUNITS OF HUMAN LUTEINIZING HORMONE

1971 ◽  
Vol 51 (1) ◽  
pp. 169-180 ◽  
Author(s):  
ANNE STOCKELL HARTREE ◽  
MARJORIE THOMAS ◽  
MARY BRAIKEVITCH ◽  
E. T. BELL ◽  
D. W. CHRISTIE ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Thyroid Stimulating Hormone ◽  
Human Chorionic Gonadotrophin ◽  
Human Thyroid ◽  
Published Data ◽  
Immunological Activity ◽  
Human Pituitary ◽  
Α Subunit ◽  
Counter Current ◽  
Counter Current Distribution

SUMMARY Separation of the subunits of human pituitary luteinizing hormone (LH) has been accomplished by dissociation of the hormone with 8 m-urea followed by chromatography on CM-cellulose at pH 5·0 or by counter-current distribution. Each procedure resulted in isolation of one subunit, the other subunit being contaminated with reassociated LH. The isolated subunits (α and β) are each of low biological activity, but when recombined in a 1:1 ratio at pH 7, reassociation occurs with restoration of full activity. The amino acid compositions of the two subunits are significantly different from each other, but the composition of the α subunit is very similar to published data on the corresponding subunit of human chorionic gonadotrophin (HCG) and human thyroid-stimulating hormone. Significant differences were observed when comparing the compositions of the β subunits of the hormones. By immunoradiometric assay using antibodies to HCG, the α subunit was as active as native LH on a weight basis, but the β subunit was significantly lower in immunological activity.

PRODUCTION OF ANTISERA AGAINST HIGHLY PURIFIED HUMAN FOLLICLE-STIMULATING HORMONE, LUTEINIZING HORMONE AND THYROID-STIMULATING HORMONE

1976 ◽  
Vol 70 (3) ◽  
pp. 335-344 ◽  
Author(s):  
J. I. THORELL ◽  
B. HOLMSTRÖM
Keyword(s):  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Thyroid Stimulating Hormone ◽  
High Titre ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Antigenic Determinants ◽  
High Avidity ◽  
Cross Reactions ◽  
Stimulating Hormone

SUMMARY Antisera were produced in rabbits against highly purified preparations of human LH (2000 or 10000 i.u./mg), human FSH (5500 i.u./mg), and human TSH (7·5 i.u./mg). Most rabbits produced antisera of high titre and high avidity. Cross-reactions were minimal between human TSH and human chorionic gonadotrophin (HCG) and between human FSH and HCG but marked between human LH and HCG. TSH and FSH also showed a constant but relatively weak cross-reaction. LH cross-reacted with FSH to a higher degree than did HCG. The avidity of the antisera was high. It was concluded that much of the lack of specificity recorded for glycoprotein antisera are effects of impure immunogens. Some of the true cross-reactions are probably explained by shared antigenic determinants of the β-subunits. Unadsorbed antisera could be used for assay of FSH and TSH in plasma from pregnant women.

scholarly journals Treatment of immature mice with gonadotrophins. The influence of mouse age on the response of ovarian alkaline phosphatase activities to gonadotrophins

1975 ◽  
Vol 152 (2) ◽  
pp. 365-372 ◽  
Author(s):  
Thomas A. Bramley
Keyword(s):  
Alkaline Phosphatase ◽  
Luteinizing Hormone ◽  
Alkaline Phosphatase Activity ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Uterine Weight ◽  
Human Pituitary ◽  
Serum Luteinizing Hormone ◽  
Stimulating Hormone

Treatment of mice aged 23–25 days with chorionic gonadotrophin induced large amounts of an ovarian alkaline phosphatase activity (phosphatase Ib) kinetically distinct from that of untreated ovaries (phosphatase I). The activities of alkaline phosphatase I and Ib varied with age in untreated mice. Phosphatase Ib appeared when serum luteinizing hormone concentrations increased (days 4–10 and days 35–45), and disappeared when concentrations were low (days 11–35). Injection of human chorionic gonadotrophin induced progressively larger amounts of phosphatase Ib activity between day 19 and day 29. However, gonadotrophin treatment failed to induce this activity on days 10–18 and 30–35. Nevertheless, during the latter period, human chorionic gonadotrophin induced especially large increases in uterine weight. Treatment at different ages with sheep luteinizing hormone plus human pituitary follicle-stimulating hormone induced a pattern of response identical with that induced by human chorionic gonadotrophin, although sheep luteinizing hormone alone was ineffective before 35 days. In contrast, human luteinizing hormone induced a response in the absence of exogenous follicle-stimulating hormone.

Report of the Medical Research Council Committee on Human Pituitary Hormones: Radioimmunoassay Kits for Human Follicle Stimulating Hormone and Luteinizing Hormone

1971 ◽  
Vol 49 (7) ◽  
pp. 685-687 ◽  
Author(s):  
C. Faiman ◽  
B. Shome
Keyword(s):  
Luteinizing Hormone ◽  
Medical Research ◽  
Research Council ◽  
Medical Research Council ◽  
Follicle Stimulating Hormone ◽  
Pituitary Hormones ◽  
Serum Samples ◽  
Human Pituitary ◽  
Stimulating Hormone

The methodology used in the development of radioimmunoassay kits for human follicle stimulating hormone (FSH) and luteinizing hormone (LH) which are ready for distribution to qualified investigators in Canada is described. Values obtained in measuring the FSH and LH content of serum samples are virtually identical to those obtained with parallel determinations using previously described radioimmunoassay techniques.

Therapeutic use of gonadotrophins and clomiphene

1969 ◽  
Vol 7 (9) ◽  
pp. 33-35
Keyword(s):  
Pregnant Women ◽  
Follicle Stimulating Hormone ◽  
Therapeutic Use ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Synthetic Compound ◽  
Pituitary Glands ◽  
Post Menopausal ◽  
Stimulating Hormone

The three substances now used to stimulate the gonads in infertility are human follicle stimulating hormone (HFSH) obtained mainly from post-menopausal urine, but also from human pituitary glands, human chorionic gonadotrophin (HCG) extracted from the urine of pregnant women, and clomiphene (Clomid - Merrell), a synthetic compound which we reviewed in 1967.1

ANTIGONADOTROPHIC PROFILE OF ANTISERA AGAINST HUMAN GONADOTROPHIN PREPARATIONS

1971 ◽  
Vol 67 (2) ◽  
pp. 262-276 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Luteinizing Hormone ◽  
Total Dose ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Immunological Activity ◽  
Human Menopausal Gonadotrophin ◽  
Stimulating Hormone

ABSTRACT Forty-two antisera were prepared in rabbits against human chorionic gonadotrophin (HCG), human hypophysial gonadotrophin (HHG), human urinary luteinizing hormone (LH) and human menopausal gonadotrophin (HMG) preparations. The gonadotrophic profiles of the antigens were previously characterized by bioassay, immunoassay and bioimmunoassay methods. The 25 most potent antisera were tested in statistically valid bioassays for their HCG and follicle stimulating hormone (FSH) neutralizing activities as well as for their neutralizing potencies against the FSH-like activity present in HCG preparations. The anti-HCG/anti-FSH ratios of the anti-HCG sera tested varied between 6.2 and > 254, while those of the anti-HHG, anti-LH and anti-HMG sera were close to 2. It was found that the total dose of immunological activity (anti-HCG neutralizing and anti-FSH neutralizing potency) rather than that of the biological activity administered to the rabbits was decisive for obtaining antisera with high anti-HCG and anti-FSH titers. Immunization with a highly purified HCG preparation (> 17 000 IU/mg) resulted in antisera exhibiting lower anti-HCG/anti-FSH ratios than did immunization with partially purified preparations. A highly purified urinary LH preparation which did not contain any detectable FSH activity gave rise to antisera exhibiting anti-HCG/anti-FSH ratios of approximately 2.0. These highly purified HCG and LH preparations were shown previously to possess high anti-FSH neutralizing potencies (Petrusz et al. 1971b). Booster injections did not change significantly the quality or the titer of the antigonadotrophic sera studied. The HCG neutralizing potency of anti-HCG sera was approximately 3 times higher when assayed against a highly purified HCG preparation (> 17 000 IU/mg) as compared to potency estimates obtained against the laboratory standard of HCG (about 2000 IU/mg). It is suggested that consideration should be given to the establishment of standard preparations of antigonadotrophic sera. It is concluded that bioimmunoassays are more suitably than conventional bioassay methods for the assessment of the antigenic purity of human gonadotrophin preparations.

OBSERVATIONS ON THE ASSAY OF HUMAN URINARY FOLLICLE-STIMULATING HORMONE BY THE AUGMENTATION TEST IN MICE

1966 ◽  
Vol 35 (2) ◽  
pp. 199-206 ◽  
Author(s):  
P. S. BROWN ◽  
M. WELLS
Keyword(s):  
No Value ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
High Potency ◽  
Human Pituitary ◽  
Ovarian Weight ◽  
C3h Mice ◽  
C57bl Mice ◽  
Stimulating Hormone

SUMMARY The follicle-stimulating hormone (FSH) content of urinary gonadotrophic extracts was assayed by its effect on the ovarian weight of immature mice when given in conjunction with 40 i.u. human chorionic gonadotrophin. About three-quarters of all routine assays gave values of λ between 0·15 and 0·30. Precision was slightly increased when the material was given in three rather than in five injections. Correction of ovarian weight for body weight was either invalid or of no value in reducing variance. Removal of between-litter variance increased precision considerably. Mice of three randomly bred colonies were all satisfactory, and inbred C57BL mice were also suitable for the assay. C3H mice were less sensitive. The efficiency of different methods of extracting FSH from urine was examined. The method of Johnsen (1958) using precipitation with tannic acid was considered the most satisfactory and gave extracts of high potency and low bulk. Limited experiments in which purified human pituitary FSH was assayed with and without added luteinizing hormone, gave results compatible with the assumption that the method is specific for FSH.

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