RELATIONSHIPS BETWEEN PHYSICO-CHEMICAL, IMMUNOLOGICAL AND BIOLOGICAL PROPERTIES OF HUMAN CHORIONIC GONADOTROPHIN

1971 ◽  
Vol 67 (3) ◽  
pp. 434-444 ◽  
Author(s):  
D. L. Matthies ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Gel Filtration ◽  
First Trimester ◽  
Biological Properties ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Separate Component ◽  
Immunological Activity ◽  
Physico Chemical ◽  
Pregnancy Urine

ABSTRACT Gel filtration of pregnancy urine on Sephadex G-100 separates two molecular species related to human chorionic gonadotrophin (HCG). One is a slightly retarded fraction which possesses biological activity but little or no immunological activity. The other is a highly retarded fraction which possesses immunological activity but no detectable biological activity. The highly retarded material could not be detected in pregnancy serum nor in extracts of first trimester placentas. A nonpregnant woman injected intravenously with purified HCG which displayed only slight retardation on the column produced urine which contained the highly retarded material. Experiments are described, the results of which suggest that the highly retarded material is a separate component of urine, probably generated by the kidney.

RELATIONSHIPS BETWEEN PHYSICO-CHEMICAL IMMUNOLOGICAL AND BIOLOGICAL PROPERTIES OF HUMAN CHORIONIC GONADOTROPHIN

1971 ◽  
Vol 67 (3) ◽  
pp. 445-456 ◽  
Author(s):  
D. L. Matthies ◽  
P. Petrusz ◽  
E. Diczfalusy
Keyword(s):  
Follicle Stimulating Hormone ◽  
Gel Filtration ◽  
Biological Properties ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Gel Filtration Chromatography ◽  
Physico Chemical ◽  
Immunological Test ◽  
Stimulating Hormone

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.

COMPARISON OF BIOASSAY AND IMMUNOASSAY OF HUMAN CHORIONIC GONADOTROPHIN IN URINE

1965 ◽  
Vol 50 (3) ◽  
pp. 335-344 ◽  
Author(s):  
Rudi Borth ◽  
Michel Ferin ◽  
Annette Menzi
Keyword(s):  
Biological Activity ◽  
Total Variation ◽  
Regression Line ◽  
Haemagglutination Inhibition ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Factorial Experiment ◽  
Serological Result ◽  
Pregnancy Urine ◽  
Acceptable Agreement

ABSTRACT In 39 samples of pregnancy urine, the concentration of human chorionic gonadotrophin (HCG) was estimated biologically by the ovarian hyperaemia reaction in rats, and serologically by the passive haemagglutinationinhibition technique. The results of the bioassays varied from 3 to 150 IU/ml, those of the immunoassays from 5 to 640 IU-eq./ml, and the correlation between the two (calculated for their logarithms) accounted for only 17 per cent of the total variation (r2 = 0.169, P ≈0.01). If the biological activity were estimated from a serological result and the appropriate regression line, the fiducial interval for P = 0.05 would extend from 17 to 610 per cent of the estimate. In a factorial experiment using three anti-HCG sera, three standard and three sensitizing preparations of HCG, the sensitivity of the serological system (expressed as the endpoint concentration in IU of HCG) varied considerably between the 27 combinations of the 3 factors, but there was no interaction between the latter. From these data and those of other authors, it is concluded that immunoassays based on haemagglutination inhibition cannot replace bioassays in the estimation of HCG, as distinct from its hypothetical metabolites or other related antigens, unless specificity has been demonstrated. The well-documented reliability of serological pregnancy tests is, of course, not in dispute. Attention is drawn to the fact that »statistically significant« correlation does not guarantee analytically acceptable agreement between two methods of assay.

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.

DIFFERENCES IN THE BIOLOGICAL AND IMMUNOLOGICAL ACTIVITIES OF HUMAN CHORIONIC GONADOTROPHIN AFTER THE REMOVAL OF SIALIC ACID BY ENZYMIC HYDROLYSIS

1967 ◽  
Vol 38 (4) ◽  
pp. 395-399 ◽  
Author(s):  
W. A. BARR ◽  
J. G. COLLEE
Keyword(s):  
Biological Activity ◽  
Sialic Acid ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Enzymic Hydrolysis ◽  
Immunological Activity ◽  
Parallel Reduction ◽  
Free Sialic Acid

SUMMARY Incubation of a partially purified preparation of human chorionic gonadotrophin (HCG) with neuraminidase resulted in release of free sialic acid from the substrate. The biological activity of the HCG was considerably reduced after incubation with the enzyme, but a parallel reduction in immunological activity was not observed.

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.

SUBUNITS OF HUMAN CHORIONIC GONADOTROPHIN: IMMUNOLOGICAL AND BIOLOGICAL STUDIES

1975 ◽  
Vol 79 (4) ◽  
pp. 749-766 ◽  
Author(s):  
S. Donini ◽  
I. D'Alessio ◽  
P. Donini
Keyword(s):  
Biological Activity ◽  
Gel Filtration ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Biologically Active ◽  
Β Subunit ◽  
Reactive Material ◽  
Biological Studies ◽  
The Cross

ABSTRACT The α and β subunits of human chorionic gonadotrophin (hCG) were prepared by incubation in 8 m urea, pH 4.5. The separation of the two subunits was obtained by DEAE-Sephadex A-25 chromatography and purification was carried out by gel filtration on Sephadex G-100. The β subunit obtained was biologically active and was therefore further purified by affinity chromatography using as immuno-adsorbent the α antibodies coupled to Sepharose 4B. The β subunit so purified showed a biological activity less than 1 IU/mg. The immunological and biological properties of the hCG subunits have been studied. It was found that the anti hCG β serum can discriminate between hCG and hLH and that in the 125I-hCG + anti-β serum radioimmunoassay, the cross-reactivity of pituitary hLH was lower than that of urinary hLH. Moreover, it was observed that the less purified was the urinary LH preparation, the higher was the cross-reactivity. Therefore we considered the hypothesis that during the purification of human menopausal gonadotrophin (hMG) some LH subunits or smaller immunoreactive fragments could have been discarded with the waste fractions. In order to test the validity of this hypophysis, all the protein fractions obtained during the purification of the hMG were gel-filtered on Sephadex G-100. The immunoreactivity of the effluents from the gel filtration was tested by hCG, hCG-β, hCG-α and hLH radioimmunoassays. While the α reactive material was found in some fractions as a peak having the same Ve/Vo value as hCG-α, the β reactive material present in the crude hMG fractions was not observed in other fractions. The cross-reactivity with the anti β serum was very low and was found in the LH region of the gel chromatogram. Furthermore, the neutralization of the biological activity of hCG and of urinary and pituitary LH by the anti hCG β serum was studied by incubating a fixed amount of the three hormones with increasing volumes of antiserum and measuring the LH activity after incubation by the OAAD test. It was observed that the anti hCG β serum inhibits hCG more than urinary or pituitary LH.

THE IMMUNOLOGICAL AND BIOLOGICAL ACTIVITY OF HUMAN CHORIONIC GONADOTROPHIN IN URINE

1964 ◽  
Vol 46 (4) ◽  
pp. 632-638 ◽  
Author(s):  
Bruce Hobson ◽  
Leif Wide
Keyword(s):  
Biological Activity ◽  
Pregnant Women ◽  
Hydatidiform Mole ◽  
Haemagglutination Inhibition ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Immunological Activity ◽  
Almost All ◽  
Inhibition Reaction ◽  
Rat Seminal Vesicle

ABSTRACT When assayed against the International Standard for HCG the biological activity, as measured by the rat seminal vesicle method, of urines from women collected during the second half of pregnancy is lower than the immunological activity (haemagglutination inhibition reaction). Almost 100 % of the immunological and biological HCG activities were recovered from the acetone precipitates of such urines. A kaolin extract of these urines produced a partial separation of the immunological activity. About half of the immunological activity and almost all of the biological activity was recovered in the concentrate. In the supernatant, left after kaolin extraction, an immunologically active biologically inactive »HCG« was found. A urine from a woman with a hydatidiform mole was assayed by both methods. The biological and immunological activities of this urine were almost unity and the ratio of the 2 activities remained unaltered in the acetone precipitate and the kaolin concentrate made from an aliquot of this urine. The kaolin supernatant contained equal and measurable amounts of the biological and immunological activities. In conclusion the method used to concentrate the urine of pregnant women will affect the ratio between the biological activity and the immunological HCG activity.

Studies of the human chorionic gonadotrophin-like substance of human pituitary glands and its significance

1983 ◽  
Vol 96 (1) ◽  
pp. 115-126 ◽  
Author(s):  
Anne Stockell Hartree ◽  
Ratna C. Shownkeen ◽  
V. C. Stevens ◽  
Shuji Matsuura ◽  
Masanobu Ohashi ◽  
...  
Keyword(s):  
Gel Filtration ◽  
Molecular Size ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Partial Purification ◽  
Human Pituitary ◽  
Human Pituitary Gland ◽  
Diethylaminoethyl Cellulose ◽  
Pituitary Glands ◽  
Pregnancy Urine

Radioimmunoassays utilizing antisera specific for the carboxyl-terminal portion of human chorionic gonadotrophin (hCG) β-subunit were used to measure the concentration in human pituitary extracts of an immunoactive hCG factor (hCG′) which was different from human LH (hLH). The content of hCG′ from different human pituitary pools collected between 1966 and 1979 was relatively constant at 0·5–1·1 μg per gland. The hCG′ concentrations observed in acetone-dried powder of individual human pituitary glands (0·4–26 ng/mg) were close to those reported for full-term placental powder. After separation and partial purification of human pituitary glycoprotein hormones, pituitary hCG′ was found mainly in the crude human FSH (hFSH) fraction. It was separated from hFSH by diethylaminoethyl–cellulose chromatography at pH 7 and by gel filtration on Sephadex G-100. On gel filtration its molecular size was larger than that of hLH or hFSH and it was strongly bound to Concanavalin A–Sepharose. The most active preparations of pituitary hCG' obtained by these procedures were approximately 5 per cent as potent by specific radioimmunoassay as hCG purified from pregnancy urine. Although the hCG′ content in individual pituitary glands was more variable than the hLH content, on average pituitary hCG′ was estimated to be around 25- to 50-fold less than the content of hLH, hFSH or human TSH in the human pituitary gland.

Biological Function and Chemistry of Endothelin. A Review

1999 ◽  
Vol 64 (8) ◽  
pp. 1211-1252 ◽  
Author(s):  
Jan Hlaváček ◽  
Renáta Marcová
Keyword(s):  
Biological Activity ◽  
Amino Acid ◽  
Biological Function ◽  
Biological Properties ◽  
Structural Basis ◽  
Amino Acid Residues ◽  
Snake Venoms ◽  
Vasoactive Peptides ◽  
Physico Chemical ◽  
Endothelin A

The first part of this review deals with the biosynthesis and a biological function of strongly vasoactive peptides named endothelins (ETs) including vasoactive intestinal contractor. Where it was useful, snake venoms sarafotoxins which are structural endothelin derivatives, were also mentioned. In the second part, an attention is paid to structural basis of the ETs biological activity, with respect to alterations of amino acid residues in the parent peptides modifying the conformation and consequently the physico-chemical and biological properties in corresponding ETs analogs. Special attention is focussed on the area of ETs receptors and their interaction with peptide and non peptide agonists and antagonists, important in designing selective inhibitors of ETs receptors potentially applicable as drugs in a medicine. A review with 182 references.

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