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.

IMMUNOLOGICAL CROSS REACTION BETWEEN HUMAN CHORIONIC GONADOTROPHIN AND HUMAN PITUITARY GONADOTROPHINS

1966 ◽  
Vol 53 (3) ◽  
pp. 420-428 ◽  
Author(s):  
C. Robyn ◽  
P. O. Hubinont ◽  
E. Diczfalusy
Keyword(s):  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Specific Antisera ◽  
Human Menopausal Gonadotrophin ◽  
Immunological Cross Reaction

ABSTRACT Immunologically mono-specific antisera prepared against human chorionic gonadotrophin (HCG) preparations completely neutralized in vitro as well as in vivo the luteinizing hormone (LH) and also the follicle-stimulating hormone (FSH) activity of both human hypophyseal gonadotrophin (HHG) and human menopausal gonadotrophin (HMG) preparations.

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.

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.

ANTIGONADOTROPHIC PROFILE OF ANTISERA AGAINST HUMAN GONADOTROPHIN PREPARATIONS

1971 ◽  
Vol 67 (2) ◽  
pp. 249-261 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Luteinizing Hormone ◽  
Biological Activities ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Cross Reaction ◽  
Chemical Methods ◽  
Physico Chemical ◽  
Human Menopausal Gonadotrophin

ABSTRACT Human chorionic gonadotrophin (HCG), human menopausal gonadotrophin (HMG) and human hypophysial gonadotrophin (HHG) preparations were assayed by two bioassay methods for their HCG or luteinizing hormone (LH) and follicle stimulating hormone (FSH) activities and by two bioimmunoassay techniques for their anti-HCG neutralizing and anti-FSH neutralizing potencies. The immunological activities measured by bioimmunoassays were expressed in anti-anti-units (AAU) according to Petrusz et al. (1971a). Seven of the HCG preparations tested showed a good correlation between their HCG, FSH-like and anti-FSH neutralizing activities. An increase of 1000 IU/mg in the specific HCG activity was usually associated with an increase of 1 IU equivalent of FSH-like activity and with an increase of 100 AAU of the anti-FSH neutralizing potency. Four HCG preparations did not contain any detectable FSH-like activity; also these preparations neutralized high amounts of anti-FSH antibodies. Two highly purified HCG preparations possessed a much lower anti-FSH neutralizing potency than was expected on the basis of their specific HCG activities. These observations seem to indicate that some of the components responsible for the cross-reaction with FSH can be removed from HCG preparations by physico-chemical methods. The anti-HCG neutralizing potencies of partially purified HCG preparations agreed fairly well with their biological activities. In highly purified HCG preparations the biological activity exceeded 2 to 5 times the anti-HCG neutralizing potency. The anti-FSH and anti-HCG neutralizing potencies of HMG preparations having FSH/LH ratios close to unity were very similar to their biological FSH and LH activities, respectively. One LH preparation, purified from HMG and lacking detectable biological FSH activity, exhibited a high anti-FSH neutralizing potency. The anti-HCG neutralizing and anti-FSH neutralizing activities of the two HHG preparations tested were some 3 to 5 times more than their biological LH and FSH activities, respectively. It is concluded that the biological purity of human gonadotrophin preparations has little relevance to their immunological purity.

THE ACTION OF FOLLICLE STIMULATING HORMONE AND OF HUMAN CHORIONIC GONADOTROPHIN UPON STEROID SYNTHESIS BY RABBIT OVARIAN TISSUES IN VITRO

1964 ◽  
Vol 47 (2) ◽  
pp. 293-305 ◽  
Author(s):  
Denis Gospodarowicz
Keyword(s):  
Metabolic Pathways ◽  
Specific Activity ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Steroid Synthesis ◽  
Site Of Action ◽  
Metabolic Sequence ◽  
Stimulating Hormone

ABSTRACT Rabbit ovarian follicles have been incubated with sodium acetate-14C with and without the presence of gonadotrophin. The major radioactive metabolites found were dehydroepiandrosterone (DHEA), androst-5-ene-3β,17β-diol, testosterone, and 17α-oestradiol. Lesser amounts of radioactivity were found in pregnenolone, progesterone, androstenedione, and oestrone. The results are interpreted as evidence that two metabolic pathways exist in the follicle; the major one giving rise to androgens and oestrogens from Δ5-3β-hydroxysteroids while the other pathway involves pregnenolone and progesterone as intermediates. Follicle stimulating hormone (FSH) and human chorionic gonadotrophin (HCG) both increased the incorporation of acetate-14C into steroids and decreased the specific activity of cholesterol. The site of action of the gonadotrophins in the metabolic sequence is discussed.

Twin Pregnancies Following Induction of Ovulation: A Literature Review

1982 ◽  
Vol 31 (3-4) ◽  
pp. 247-253 ◽  
Author(s):  
John A. Lamont
Keyword(s):  
Literature Review ◽  
Clomiphene Citrate ◽  
Vital Statistics ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Multiple Pregnancies ◽  
Human Pituitary ◽  
True Incidence ◽  
Induction Of Ovulation ◽  
Human Menopausal Gonadotrophin

A literature review of the occurrence of multiple pregnancies associated with artificial induction of ovulation is reported. This report considers three treatment schedules: (1) clomiphene citrate; (2) human pituitary gonadotrophin with human chorionic gonadotrophin; and (3) human menopausal gonadotrophin with human chorionic gonadotrophin. The majority of the increase in twinning is related to hyperstimulation of the ovary by these medications, resulting in dizygotic twinning. The true incidence of twin pregnancy cannot be calculated because the vital statistics of all nations report live birth rates. Increased rates of fetal wastage, late abortion and prematurity associated with the occurrence of multiple pregnancies are overlooked by these statistics. The increased incidence of twinning appears to be related to the type and dosage of medication used, and the patient's underlying problem.

BIOASSAY OF ANTIGONADOTROPHIC SERA

1968 ◽  
Vol 59 (2) ◽  
pp. 261-276 ◽  
Author(s):  
C. Robyn ◽  
E. Diczfalusy
Keyword(s):  
Reproductive Organs ◽  
Human Chorionic Gonadotrophin ◽  
Male Rats ◽  
Ventral Prostate ◽  
International Standard ◽  
Immature Male ◽  
Antigenic Properties ◽  
Human Menopausal Gonadotrophin ◽  
Reference Preparation ◽  
Second International

ABSTRACT Methods are described for the bioassay of the human chorionic gonadotrophin (HCG) – and luteinising hormone (LH) neutralising potencies of antigonadotrophic sera. The methods are based on the increase in weight of the accessory reproductive organs of intact immature male rats and on the increase in weight of the ventral prostate of hypophysectomised immature rats. The antigonadotrophic sera were obtained following immunisation of rabbits with HCG, human menopausal gonadotrophin (HMG) and human hypophysial gonadotrophin (HHG) preparations. These sera were then assayed against the laboratory standard and the International Standard preparations of HCG, as well as against a highly purified HCG preparation. The antigonadotrophic potencies of the various antisera showed a close agreement, when assayed in intact or hypophysectomised animals against the different HCG preparations. When anti-HCG sera were assayed in intact or hypophysectomised animals against the laboratory standard of HMG, the antigonadotrophic potencies were approximately three times higher than those obtained against HCG preparations. In an attempt to account for this discrepancy, the laboratory standard of HCG was assayed against the laboratory standard of HMG and the Second International Standard preparation of HCG was assayed against the Second International Reference preparation of HMG in both assay systems used. The potency of 1.0 IU of HCG corresponded to that of 2.5 to 2.9 IU of LH. If this difference is taken into consideration, the discrepancy in antigonadotrophic titers disappears. However, when anti-HCG and anti-HHG sera were assayed in intact or hypophysectomised animals against an HHG preparation, the antigonadotrophic potencies were approximately ten times lower than those obtained when the antisera were tested against HCG preparations and ten to thirty times lower than those obtained in the assays conducted against the laboratory standard of HMG. Since these discrepancies in antigonadotrophic titers cannot be explained by differences in the gonadotrophic potency of the preparations used, it is concluded, that major differences exist in the antigenic properties of human gonadotrophins of pituitary – and urinary origin.

Sign in / Sign up

Export Citation Format

Share Document