STRAIN VARIATION IN THE ASSAY OF FOLLICLE-STIMULATING HORMONE BY THE OVARIAN AUGMENTATION TEST IN MICE

1970 ◽  
Vol 63 (2) ◽  
pp. 275-282
Author(s):  
E. T. Bell ◽  
D. W. Christie
Keyword(s):  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
National Institutes Of Health ◽  
Strain Variation ◽  
Ovarian Weight ◽  
Human Menopausal Gonadotrophin ◽  
Reference Preparation ◽  
Very High ◽  
Stimulating Hormone

ABSTRACT Assays of follicle-stimulating hormone from the National Institutes of Health (NIH-FSH-S4) and the Second International Reference Preparation for Human Menopausal Gonadotrophin (IRP-HMG) have been conducted by the mouse ovarian augmentation test in animals of nine strains from five mouse breeders. Two groups of 50 mice from each colony were used to assay NIH-FSH and the Second IRP-HMG. In the experiment with NIH-FSH dosages of 37.5 to 300.0 μg were given together with 40 IU human chorionic gonadotrophin (HCG) in three or five sc injections over three days. When the Second IRP-HMG was assayed dosages of 0.19 to 1.5 IU were administered in five injections with 20 or 40 IU HCG. Little or no ovarian weight increase occurred following NIH-FSH in six out of nine colonies. In the remaining three the index of precision (λ) was very high. Following administration of the Second IRP-HMG a greater increase in ovarian weight occurred but a satisfactory slope was noted in only three colonies. The λ figures were generally lower than with NIH-FSH. It is concluded, that under the conditions used, six out of the nine colonies were not suitable for the assay of FSH by the ovarian augmentation test. Further work would be required to study the reliability criteria of the assay in the remaining three colonies.

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.

FOLLICLE STIMULATING HORMONE-LIKE ACTIVITY IN HUMAN CHORIONIC GONADOTROPHIN PREPARATIONS

1969 ◽  
Vol 60 (1) ◽  
pp. 137-156 ◽  
Author(s):  
C. Robyn ◽  
P. Petrusz ◽  
E. Diczfalusy
Keyword(s):  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Female Rats ◽  
Immature Female ◽  
Ovarian Weight ◽  
Wide Range ◽  
Immature Rats ◽  
Stimulation Of ◽  
Stimulating Hormone

ABSTRACT The follicle stimulating hormone (FSH)-like activity of human chorionic gonadotrophin (HCG) preparations was assayed by the method based on the ovarian weight augmentation in intact immature rats. The potencies ranged from 4.8 to 7.4 IU equivalents of FSH per mg. The FSH-like potency of the Second International Standard Preparation of HCG was 8.5 IU per vial. However, when in intact immature rats the ovarian weight response to HCG preparations was compared at a wide range of doses (40 to 51 200 IU) to that obtained with a human menopausal gonadotrophin (HMG) preparation (0.5 to 128 IU of FSH) in the presence of 40 IU of HCG, significant differences were found. The assays conducted in hypophysectomised immature female rats were invalid, because of lack of parallelism. Antisera were prepared by immunising rabbits with HCG and human hypophysial gonadotrophin (HHG) preparations and the antigonadotrophin profiles (HCG-, FSH- and FSH-like neutralising potencies) of these antisera were established by the use of statistically valid bioassay procedures. The anti-HCG and anti-HHG sera neutralised the FSH activity of HMG preparations as well as the FSH-like activity of HCG preparations. However, 3 to 175 times more antiserum was required to neutralise the equivalent of 1.0 IU of FSH-like activity present in HCG than expected on the basis of the anti-FSH potency of the antisera. On the other hand, there was a high degree of correlation between the neutralising potencies of the antisera when tested against the FSH-like activity and the HCG activity of various HCG preparations. When the FSH-like activity of an HCG preparation was quantitatively neutralised with an anti-HCG serum, some 30 per cent of the HCG activity remained unneutralised, as evidenced by repeated bioassays. Although at least 2000 IU of this »FSH-free« HCG was administered to groups of intact as well as hypophysectomised immature female rats, this high dose of HCG did not induce an increase in ovarian weight beyond that elicited by 40 IU of untreated HCG. Histological examination of the ovaries indicated lack of follicle stimulation in the hypophysectomised, but not in the intact immature animals. There was an excessive stimulation of the interstitial cells in both types of animals. The data indicate that the FSH-like activity of HCG preparations is neither due to a contamination by FSH of pituitary origin, nor is it an evenly distributed intrinsic property of the HCG molecules. It is also concluded that the gonadotrophic activity of biologically pure HCG in immature hypophysectomised female rats consists of a specific stimulation of the interstitial cell apparatus. Such HCG preparations do not induce any follicle stimulation, not even when administered in excessive doses.

FOLLICLE STIMULATING HORMONE IN THE URINE OF PREGNANT WOMEN

1957 ◽  
Vol 16 (1) ◽  
pp. 107-113 ◽  
Author(s):  
W. R. BUTT ◽  
A. C. CROOKE ◽  
JOYCE D. INGRAM ◽  
BRENDA P. ROUND
Keyword(s):  
Benzoic Acid ◽  
Pregnant Women ◽  
Follicle Stimulating Hormone ◽  
Normal Pregnancy ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Ovarian Weight ◽  
Preliminary Results ◽  
Stimulating Hormone

SUMMARY 1. Follicle stimulating hormone (FSH) has been obtained from the urine of pregnant women. 2. It was prepared by adsorption on kaolin from urine which had been treated with benzoic acid to remove excess human chorionic gonadotrophin (HCG) and was assayed by the procedure which depends on the increase in ovarian weight of immature mice treated simultaneously with HCG. 3. Preliminary results are given for the assay of FSH in normal pregnancy.

BIOASSAY OF ANTIGONADOTROPHIC SERA

1968 ◽  
Vol 59 (2) ◽  
pp. 277-297 ◽  
Author(s):  
C. Robyn ◽  
E. Diczfalusy
Keyword(s):  
Specific Activity ◽  
Follicle Stimulating Hormone ◽  
High Specific Activity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Antigenic Properties ◽  
Human Menopausal Gonadotrophin ◽  
Reference Preparation ◽  
Quantitative Manner

ABSTRACT Methods are described for the bioassay of the human follicle stimulating hormone (FSH) neutralising potency of antigonadotrophic sera. The methods are based on a modified ovarian weight augmentation test using human chorionic gonadotrophin (HCG) or luteinising hormone (LH) of ovine origin for augmentation. The antigonadotrophic sera were obtained following immunisation of rabbits with HCG, human menopausal gonadotrophin (HMG) and human hypophysial gonadotrophin (HHG) preparations. The FSH neutralising potencies of these antisera were assayed against laboratory standard preparations of HMG and HHG and against the Second International Reference Preparation of HMG. When HCG was used for augmentation, the FSH neutralising potency of antisera depended on the sequence in which HCG, HMG and antiserum were combined. When HCG was mixed with the antiserum prior to the addition of HMG, this resulted in a significant decrease in the FSH neutralising potency. When HCG was injected separately from the HMG-antiserum complex, the FSH neutralising potency increased. However, the FSH neutralising potency of all antisera was significantly higher when LH of ovine origin, rather than HCG was used for augmentation. Anti-HCG sera exhibited a considerable FSH neutralising potency, even when prepared by immunisation with HCG preparations of high specific activity. These high FSH neutralising potencies were in contrast to the low FSH activity of the HCG preparations used for immunisation. Anti-HMG sera possessed little, if any, FSH neutralising potency. These poor FSH neutralising potencies were in contrast to the high FSH activity of the HMG preparations used for immunisation. The FSH neutralising potency of an anti-HHG serum was at least 5 times higher when assayed against HMG, than when assayed against HHG. The data presented indicate that HCG preparations extensively compete with FSH preparations for antibodies neutralising FSH activity. This suggests that there is a cross reaction between HCG and FSH. The data also indicate, that there are significant differences in the antigenic properties of human pituitary and urinary gonadotrophins. It is concluded, that the establishment of specificity of immunoassay methods for human gonadotrophins cannot be based exclusively on immunological evidence. Also, the absorption procedures used to improve the specificity of antigens and antisera are of limited value, unless carried out in a strictly quantitative manner following the establishment of the profile of the gonadotrophic and antigonadotrophic activities present.

THE ASSAY OF GONADOTROPHIN FROM URINE OF NON-PREGNANT HUMAN SUBJECTS

1955 ◽  
Vol 13 (1) ◽  
pp. 59-64 ◽  
Author(s):  
P. S. BROWN
Keyword(s):  
Human Subjects ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Uterine Weight ◽  
Ovarian Weight ◽  
Stimulating Hormone

SUMMARY Two convenient bioassays of urinary gonadotrophins, using immature mice, are described. The first is based upon the initial doubling of uterine weight. The second, using the ovarian weight response, attempts to increase specificity to follicle stimulating hormone by priming with human chorionic gonadotrophin. The usefulness of both methods is discussed, and the influence of non-specific impurities during the assay of urinary extracts is stressed.

STUDIES ON THE MOUSE UTERINE AUGMENTATION TEST FOR THE ASSAY OF FOLLICLE-STIMULATING HORMONE IN FOUR MOUSE COLONIES

1971 ◽  
Vol 68 (4) ◽  
pp. 715-724
Author(s):  
D. W. Christie ◽  
E. T. Bell ◽  
D. Harrison
Keyword(s):  
Dose Response ◽  
Regression Coefficient ◽  
Follicle Stimulating Hormone ◽  
Human Chorionic Gonadotrophin ◽  
National Institutes Of Health ◽  
Strain Variation ◽  
Linear Regression Coefficient ◽  
Response Curves ◽  
Different Doses ◽  
Stimulating Hormone

ABSTRACT Assays of follicle-stimulating hormone from the National Institutes of Health (NIH-FSH-S4) have been conducted using the uterine augmentation test in four strains of mice. Dose response curves were determined for each strain using dosages of NIH-FSH ranging from 4–64 μg together with 0.1 IU human chorionic gonadotrophin (HCG) in five sc injections given over three days. The specificity of the assay was investigated in each strain by adding 2, 10 and 50 μg luteinizing hormone (NIH-LH-S6) to each of three doses of NIH-FSH, 6, 12 and 24 μg, forming a complete three factor experimental design; each mouse also received 0.1 IU HCG. Five mice from each strain were employed at each of the 64 treatment combinations. Values for b and λ in the dose response studies were acceptable for quantitative work but all strains were relatively insensitive. In the specificity studies the analysis of variance showed that a marked strain variation existed in the magnitude of the response to NIH-FSH and to NIH-LH while one strain showed a significant interaction between these two hormones. Regression analysis was carried out and the influence of different doses of NIH-LH on the linear regression coefficient was examined. In general it was considered that, under the conditions employed, none of the four strains would prove useful in the uterine augmentation test.

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

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