ANTIGONADOTROPHIC PROFILE OF ANTISERA AGAINST HUMAN GONADOTROPHIN PREPARATIONS

1971 ◽  
Vol 67 (2) ◽  
pp. 249-261 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy

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.

1971 ◽  
Vol 67 (2) ◽  
pp. 262-276 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy

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.


1966 ◽  
Vol 36 (1) ◽  
pp. 23-28 ◽  
Author(s):  
PACHARA VISUTAKUL ◽  
E. T. BELL ◽  
J. A. LORAINE ◽  
R. B. FISHER

SUMMARY Human chorionic gonadotrophin (HCG), pregnant mare serum gonadotrophin (PMSG) and human menopausal gonadotrophin (HMG) were incubated with varying concentrations of urea at different temperatures for different times. The luteinizing hormone (LH) activity of HCG was progressively destroyed with increasing concentrations of urea. The degree of inactivation was greater at higher temperatures but the time of incubation did not affect the results. The follicle-stimulating activity of PMSG was reduced at high urea concentrations; the time of incubation was without effect. Under the experimental conditions used the LH activity of HMG was generally destroyed to a greater extent than its follicle-stimulating hormone activity.


1966 ◽  
Vol 53 (3) ◽  
pp. 420-428 ◽  
Author(s):  
C. Robyn ◽  
P. O. Hubinont ◽  
E. Diczfalusy

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.


1964 ◽  
Vol 46 (2) ◽  
pp. 317-330 ◽  
Author(s):  
Savitri K. Shahani ◽  
Shanta Savur Rao

ABSTRACT Immunological investigations with human chorionic gonadotrophin (HCG) were carried out in order to characterize the antigens of HCG. Attempts were also made to find out whether HCG has antigens common to those of human luteinizing hormone (LH) and human follicle stimulating hormone (FSH) and also to ovine, bovine and porcine LH and pregnant mare serum gonadotrophin (PMSG). The results of the immunoelectrophoretic analysis carried out have indicated that one of the three antigens of HCG seems to be immunochemically similar to the antigen present in human LH. HCG was not observed to have any antigens in common with ovine, bovine and porcine LH and PMSG as revealed by the tests carried out with antiserum to HCG. The combining power and the biological activity of the antigen common to human LH and HCG were not completely destroyed by heating the hormones at 100° C for 30 minutes. These were, however, destroyed by treating the respective hormones with 30 per cent hydrogen peroxide. Haemorrhagic follicles were observed when human FSH was injected into immature mice together with human LH. Such haemorrhagic follicles were also observed in some of the mice injected simultaneously with heated LH or HCG along with human FSH. The significance of these observations are discussed.


1962 ◽  
Vol 24 (2) ◽  
pp. 153-158 ◽  
Author(s):  
H. SCHMIDT-ELMENDORFF ◽  
J. A. LORAINE ◽  
E. T. BELL

SUMMARY The luteinizing hormone (LH), follicle-stimulating hormone (FSH) and 'total gonadotrophic' activities of various hormones have been studied following incubation with 6 m urea at 40° c for 24 hr. LH activity was estimated by the ovarian ascorbic acid depletion test in rats, FSH activity by the augmentation test in mice, and 'total gonadotrophic' activity by the mouse uterus test. Following incubation with 6 m urea the LH activities of NIH—LH, NIH—FSH, human chorionic gonadotrophin and Pergonal were almost completely destroyed, while the LH activity of pregnant mares' serum gonadotrophin (PMSG) was reduced to a smaller extent. The FSH activity of NIH—FSH was little affected by this form of treatment, but in the case of Pergonal a considerable reduction of FSH activity occurred. The 'total gonadotrophic' activity of NIH—FSH, PMSG and Pergonal was reduced after incubation with 6 m urea, the degree of inactivation being greatest in the case of Pergonal. After control incubations with water no appreciable loss of biological activity was observed with any hormone other than Pergonal.


1983 ◽  
Vol 99 (2) ◽  
pp. 199-209 ◽  
Author(s):  
Valerie Urwin

Heterologous double-antibody radioimmunoassays were developed for the measurement of FSH and LH concentrations in the serum of both horses and donkeys. The FSH assay employed a rabbit anti-ovine FSH serum which showed a complete lack of cross-reaction with equine chorionic gonadotrophin (eCG) and negligible cross-reaction with equine LH. The LH assay utilized an antiserum raised against highly purified eCG. This similarly showed negligible cross-reaction with equine FSH but its high cross-reactivity with eCG prevented the measurement of equine LH concentrations in serum when eCG was also present. In both assays serial dilutions of horse and donkey serum were parallel to the standard. The assays were used to monitor changes in serum concentrations of FSH and LH during the first 100 days of pregnancy in pony mares and jenny donkeys. In both species during pregnancy LH levels reached a peak 1–2 days after ovulation. They then decreased rapidly to baseline levels where they remained until days 35–40 when the commencement of eCG production prevented their further measurement. Serum FSH concentrations, on the other hand, continued to fluctuate markedly throughout the first 100 days of pregnancy in both the ponies and donkeys. Pronounced surges in FSH levels occurred at regular intervals in some animals but the pattern of release was quite irregular in the others. The results of this study support the concept that it is continued pituitary FSH release, not eCG secretion, which is responsible for stimulating the secondary follicles which develop during early equine pregnancy. However, it appears likely that it is the LH-like activity of eCG which causes the subsequent ovulation and/or luteinization of these secondary follicles to produce accessory corpora lutea.


1976 ◽  
Vol 70 (3) ◽  
pp. 335-344 ◽  
Author(s):  
J. I. THORELL ◽  
B. HOLMSTRÖM

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.


2017 ◽  
Vol 23 (3) ◽  
pp. 121-125 ◽  
Author(s):  
G. A. Filipescu ◽  
Oana Alina Solomon ◽  
Nicoleta Clim ◽  
Amelia Milulescu ◽  
Andreea Gratiana Boiangiu ◽  
...  

AbstractMolar pregnancies results from a tainted fertilization process. Trophoblastic thyroidian hyper function is an unusual complication of a molar pregnancy. The degree of thyroid stimulation and the severity of clinical hyperthyroidism is directly proportional to HCG concentration. Human chorionic gonadotrophin is almost identical with TSH, luteinizing hormone (LH) and follicle-stimulating hormone, this analogy in the structure will cause cross-reactivity with their receptors. Hyperthyroid status can vary from asymptomatic hyper function to thyroid storm. Dilation and curettage represents the treatment for hyperthyroidism in molar pregnancy. Awareness of this condition is important for diagnosis and treatment.


1978 ◽  
Vol 79 (3) ◽  
pp. 407-408 ◽  
Author(s):  
M. J. ELLIS ◽  
R. A. DONALD ◽  
J. H. LIVESEY

The Medical Unit, The Princess Margaret Hospital, Christchurch 2, New Zealand (Revised manuscript received 21 August 1978) The frequent clinical and research requirement for measurement of both plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) has prompted the development of a simultaneous radioimmunoassay for these two hormones. The considerable advantages of a simultaneous method include an economy of plasma volume, assay reagents, test-tubes and, more importantly, the time required for radioactive counting and performance of the assay by technical staff. The latter two factors comprise a significant proportion of radioimmunoassay operating costs. This report describes a simultaneous radioimmunoassay based on the use of 131I-labelled FSH, 125I-labelled LH, anti-FSH serum M93 6873 (a generous gift from Professor W. R. Butt, Birmingham), anti-human chorionic gonadotrophin (HCG) serum for LH measurement (Donald, 1972) and donkey anti-rabbit precipitating serum (Wellcome Reagents, U.K.) for separation of antibody-bound and free hormones. Pituitary gonadotrophin standard (LER 907)


1971 ◽  
Vol 67 (3) ◽  
pp. 445-456 ◽  
Author(s):  
D. L. Matthies ◽  
P. Petrusz ◽  
E. Diczfalusy

ABSTRACT Purified human chorionic gonadotrophin (HCG) preparations were chromatographed on Sephadex G-100 and the distribution of the hormone on the chromatogram was studied by an immunological test and by bioassay. Each preparation studied showed evidence of inhomogeneity in terms of the degree to which it was retarded on the gel bed, but the type of inhomogeneity was variable. By the use of gel filtration chromatography it was possible to separate HCG molecules devoid of detectable follicle stimulating hormone (FSH)-like activity from molecules which possessed FSH-like activity. Evidence is presented which indicates that at least some physico-chemical manipulations utilized in the purification of HCG from urine may introduce artifacts.


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