BIOLOGICAL AND IMMUNOLOGICAL CHARACTERIZATION OF ANTIGONADOTROPHIC PROFILES

1969 ◽  
Vol 62 (1_Suppl) ◽  
pp. S31-S53 ◽  
Author(s):  
C. Robyn
Keyword(s):  
Biological Activities ◽  
Follicle Stimulating Hormone ◽  
Human Chorionic Gonadotrophin ◽  
The Past ◽  
Antigenic Properties ◽  
Biological Specificity ◽  
Human Menopausal Gonadotrophin ◽  
Quantitative Basis ◽  
Complex Relationships

ABSTRACT During the past few years a series of papers (Robyn et al. 1968; Robyn & Diczfalusy 1968a,b,c; Robyn et al. 1969; Petrusz et al. (1970) were published from this laboratory on the problems of statistically valid bioassays of gonadotrophin neutralizing potencies of antisera. These bioassays were based on the principle of »simple additivity« (or rather, subtractivity) of Hormone (H) and Antiserum (AS). Experiments conducted at several levels of neutralization of H by AS, ranging from 20 to 90 %, supported the validity of the principle of »simple additivity«. The human chorionic gonadotrophin (HCG), the luteinizing hormone (LH) and the follicle stimulating hormone (FSH) neutralizing potencies of rabbit anti-HCG sera, anti-human hypophysial gonadotrophin (anti-HHG) sera and anti-human menopausal gonadotrophin (anti-HMG) sera were estimated comparatively. The establishment of such antigonadotrophic profiles of antisera has several advantages. It makes it possible to improve the biological specificity of antisera by absorption procedures carried out on a strictly quantitative basis by selecting an appropriate amount of a gonadotrophin preparation with the adequate LH:FSH ratio. Also, it makes possible to improve the biological specificity of gonadotrophin preparations by absorption procedures carried out on a quantitative basis by selecting the appropriate excess of an antiserum with the adequate anti-LH:anti-FSH ratio. Finally the characterization of the antigonadotrophic profile of antisera together with that of the gonadotrophic profile of antigens provides improved tools for the study of the very complex relationships between the biological activities and the antigenic properties of gonadotrophic molecules. It is hoped, therefore, that the use of these methods will be of assistance in improving the specificity of current immunoassays for gonadotrophins.

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.

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.

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.

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.

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.

THE EFFECT OF UREA ON THE BIOLOGICAL ACTIVITY OF GONADOTROPHINS OF PLACENTAL, ENDOMETRIAL AND URINARY ORIGIN

1966 ◽  
Vol 36 (1) ◽  
pp. 23-28 ◽  
Author(s):  
PACHARA VISUTAKUL ◽  
E. T. BELL ◽  
J. A. LORAINE ◽  
R. B. FISHER
Keyword(s):  
Biological Activity ◽  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Hormone Activity ◽  
Experimental Conditions ◽  
Human Menopausal Gonadotrophin ◽  
Different Temperatures ◽  
Pregnant Mare Serum Gonadotrophin

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.

scholarly journals Genetic and Phenotypic Characterization of GII-4 Noroviruses That Circulated during 1987 to 2008

2010 ◽  
Vol 84 (18) ◽  
pp. 9595-9607 ◽  
Author(s):  
Yang Yang ◽  
Ming Xia ◽  
Ming Tan ◽  
Pengwei Huang ◽  
Weiming Zhong ◽  
...  
Keyword(s):  
Host Immunity ◽  
Phenotypic Characterization ◽  
Blood Group Antigens ◽  
Binding Assays ◽  
The Past ◽  
Antigenic Properties ◽  
Two Factors ◽  
Genetic Clusters ◽  
High Conservation

ABSTRACT The predominance and continual emergence of new variants in GII-4 noroviruses (NVs) in recent years have raised questions about the role of host immunity and histo-blood group antigens (HBGAs) in NV evolution. To address these questions, we performed a genetic and phenotypic characterization of GII-4 variants circulating in the past decade (1998 to 2008). Ninety-three GII-4 sequences were analyzed, and of them, 16 strains representing 6 genetic clusters were selected for further characterization. The HBGA binding properties were determined by both saliva- and oligosaccharide-binding assays using P particles as a model of NV capsid. The antigenic properties were also examined by enzyme immunoassay (EIA), Western blot analysis, and receptor blocking assay, using P-particle-specific antibodies from immunized mice and GII-4 virus-infected patients. Our results showed that 15 of the 16 GII-4 viruses bound to saliva of all A, B, and O secretors. Oligosaccharide binding assays yielded largely consistent results, although the binding affinities to some oligosaccharides varied among some strains. The only nonbinder had a mutation in the binding site. While antigenic variations were detected among the 16 strains, significant cross-blocking on the HBGA binding was also noted. Sequence alignment revealed high conservation of HBGA binding interfaces with some variations in adjacent regions. Taken together, our data suggested that the ability of GII-4 to recognize different secretor HBGAs persisted over the past decade, which may explain the predominance of GII-4 over other genotypes. Our data also indicated that both the host immunity and HBGAs play a role in NV evolution. While host immunity may continue driving NV for antigenic change, the functional selection by the HBGAs tends to lock the architecture of the capsid/HBGA interfaces and allows only limited variations outside the HBGA binding sites. A potential outcome of such counterselection between theses two factors in NV evolution is discussed.

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.

The Use of Advanced Analytical Techniques for Characterization of the Chemical, Physical, and Crystallographic Nature of a Surface

1973 ◽  
Vol 31 ◽  
pp. 132-133 ◽  
Author(s):  
R. E. Herfert
Keyword(s):  
Surface Characterization ◽  
Analytical Techniques ◽  
X Ray Diffraction ◽  
Depth Of Penetration ◽  
X Ray ◽  
The Past ◽  
Transmission Electron ◽  
Scanning Electron

Studies of the nature of a surface, either metallic or nonmetallic, in the past, have been limited to the instrumentation available for these measurements. In the past, optical microscopy, replica transmission electron microscopy, electron or X-ray diffraction and optical or X-ray spectroscopy have provided the means of surface characterization. Actually, some of these techniques are not purely surface; the depth of penetration may be a few thousands of an inch. Within the last five years, instrumentation has been made available which now makes it practical for use to study the outer few 100A of layers and characterize it completely from a chemical, physical, and crystallographic standpoint. The scanning electron microscope (SEM) provides a means of viewing the surface of a material in situ to magnifications as high as 250,000X.

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