Monoclonal antibodies against the free subunits of human chorionic gonadotrophin

1990 ◽  
Vol 125 (2) ◽  
pp. 301-309 ◽  
Author(s):  
P. Berger ◽  
R. Klieber ◽  
W. Panmoung ◽  
S. Madersbacher ◽  
H. Wolf ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Antigenic Determinants ◽  
Α Subunit ◽  
Α Chain ◽  
Fluid Levels ◽  
Α Subunits

ABSTRACT Discordant results on body fluid levels of human chorionic gonadotrophin (hCG) free α- and β-subunits under physiological and pathophysiological conditions, prompted us to raise a total of 260 monoclonal antibodies (MCA) against free hCG-α, free hCG-β, holo-hCG, human follicle-stimulating hormone and bovine luteinizing hormone; 153 MCA recognizing the human α-subunit and 28 reacting with hCG-β were extensively analysed for their intra- and interspecies cross-reactivity with homologous hormones, and for the compatibility of epitopes recognized by them. The immunological topography of free hCG-α and free hCG-β was resolved by these MCA, and epitope maps were designed. Six antigenic determinants on the free α-chain (α1–α6), clustered in three spatially distinct domains, and seven epitopes on the surface of free hCG-β (β1–β7), could be distinguished. Strikingly, three α-chain epitopes (α4, α5 and α6) were shared between various species, which is in contradiction to the concept of immunological species-specificity of α-subunits. Three determinants were found to be present only on the free subunits but not on holo-hCG (α6, β6 and β7), and only two determinants (β1 and β7) were hormone-specific for hCG. Based on this information, an immunoenzymometric assay for the free α-subunit of human glycoprotein hormones was established, with a sensitivity of 1·3 pg/well and a cross-reactivity with holo-hCG of less than 0·005% Thus this assay provides the basis for detecting free α-subunits in the presence of extremely high levels of holo-hormones, which may assist in elucidating the role of free α-subunits in man. Journal of Endocrinology (1990) 125, 301–309

The production of monoclonal antibodies to human chorionic gonadotrophin and its subunits

1983 ◽  
Vol 98 (3) ◽  
pp. 323-330 ◽  
Author(s):  
M. C. Stuart ◽  
P. A. Underwood ◽  
D. F. Harman ◽  
K. L. Payne ◽  
D. A. Rathjen ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Free Form ◽  
Binding Affinities ◽  
Β Subunit ◽  
Α Subunit ◽  
Specific Antisera ◽  
Hybridoma Technology ◽  
Circulating Levels

The difficulties encountered in producing highly specific antisera to human chorionic gonadotrophin (hCG) were overcome by the use of hybridoma technology. A panel of monoclonal antibodies directed toward hCG and its subunits was produced. Of the four antibodies which were fully characterized, one recognized the intact hCG molecule only, a second recognized only the free β-subunit, a third recognized only the free α-subunit and the fourth bound to the β-subunit of hCG both when it was in the free form and when it was associated with the α-subunit forming the intact hCG molecule. There was no significant cross-reaction of any of these antibodies with the pituitary glycoprotein hormones. The four antibodies had high binding affinities which should permit their use in immunoassays for measurement of circulating levels of hCG and its subunits.

Free α subunit of human chorionic gonadotrophin: molecular basis of immunologically and biologically active domains

1994 ◽  
Vol 140 (1) ◽  
pp. 145-154 ◽  
Author(s):  
S Dirnhofer ◽  
O Lechner ◽  
S Madersbacher ◽  
R Klieber ◽  
R de Leeuw ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Receptor Binding ◽  
Solid Phase ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Biologically Active ◽  
Molecular Organization ◽  
Antigenic Determinants ◽  
Α Subunit

Abstract Immunochemical studies were undertaken to identify surface-orientated epitopes of the free α subunit of human chorionic gonadotrophin (hCG-α) at the amino acid sequence level. We investigated the molecular organization of these epitopes, resolved the immunological topography in terms of spatial arrangement of antigenic domains and related structures to functions such as subunit association or receptor binding. Overlapping synthetic peptides covering the entire amino acid sequence of hCG-α, an enzymatically digested hCG-α subunit, and a reduced and alkylated hCG-α preparation were assayed in a solid-phase one-site enzyme-linked immunoassay, and in a solution-phase competitive radioimmunoassay (RIA). The antigenic topography was mapped by monoclonal antibodies (MCAs) in two-site binding assays (sandwich RIA). On the surface of hCG-α, seven different epitopes (α1–α7), arranged in four spatially distinct domains, could be distinguished: A, α1,2,4; B, α3,5; C, α6; D, α7. The peptides spanning hCG-α(13–18), hCG-α(17–22) and hCG-α(33–42) appeared to contribute to the formation of epitopes α2, α4 and α6 respectively. Since epitope α6 is present only on the free non-assembled subunit of different species, we concluded that the region hCG-α(33–42), which is evolutionarily highly conserved, represents a subunit assembly site. All but one epitope (α7) are destroyed by reducing and alkylating hCG-α. In contrast, chymotryptic digestion of hCG-α, leading to release of the heptapeptide hCG-α(41–47), did not affect epitope expression, indicating that this sequence is not involved in the formation of antigenic determinants. Addressing the biological properties of hCG-α epitopes by radioreceptor assay revealed that the three hCG-α peptides corresponding to epitopes α2, α4 and α6 did not displace radiolabelled hCG from its receptor, whereas any of the MCAs directed against determinants (α1–α5), shared by hCG and hCG-α, totally inhibited binding. Consistent with this, the antibodies neutralized the biological activity of hCG in terms of testosterone production in a mouse Leydig cell in vitro bioassay. We therefore concluded that hormone antibody-binding sites differ from those of hormone receptor binding, revealing no essential congruence of immunologically and biologically active domains. Journal of Endocrinology (1994) 140, 145–154

scholarly journals Characterization of components of Z-bands in the fibrillar flight muscle of Drosophila melanogaster.

1989 ◽  
Vol 109 (5) ◽  
pp. 2157-2167 ◽  
Author(s):  
J D Saide ◽  
S Chin-Bow ◽  
J Hogan-Sheldon ◽  
L Busquets-Turner ◽  
J O Vigoreaux ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Monoclonal Antibodies ◽  
Flight Muscle ◽  
Cross Reactivity ◽  
Antigenic Determinants ◽  
Immunogold Staining ◽  
Thick Filaments ◽  
The Matrix ◽  
Flight Muscles

Twelve monoclonal antibodies have been raised against proteins in preparations of Z-disks isolated from Drosophila melanogaster flight muscle. The monoclonal antibodies that recognized Z-band components were identified by immunofluorescence microscopy of flight muscle myofibrils. These antibodies have identified three Z-disk antigens on immunoblots of myofibrillar proteins. Monoclonal antibodies alpha:1-4 recognize a 90-100-kD protein which we identify as alpha-actinin on the basis of cross-reactivity with antibodies raised against honeybee and vertebrate alpha-actinins. Monoclonal antibodies P:1-4 bind to the high molecular mass protein, projectin, a component of connecting filaments that link the ends of thick filaments to the Z-band in insect asynchronous flight muscles. The anti-projectin antibodies also stain synchronous muscle, but, surprisingly, the epitopes here are within the A-bands, not between the A- and Z-bands, as in flight muscle. Monoclonal antibodies Z(210):1-4 recognize a 210-kD protein that has not been previously shown to be a Z-band structural component. A fourth antigen, resolved as a doublet (approximately 400/600 kD) on immunoblots of Drosophila fibrillar proteins, is detected by a cross reacting antibody, Z(400):2, raised against a protein in isolated honeybee Z-disks. On Lowicryl sections of asynchronous flight muscle, indirect immunogold staining has localized alpha-actinin and the 210-kD protein throughout the matrix of the Z-band, projectin between the Z- and A-bands, and the 400/600-kD components at the I-band/Z-band junction. Drosophila alpha-actinin, projectin, and the 400/600-kD components share some antigenic determinants with corresponding honeybee proteins, but no honeybee protein interacts with any of the Z(210) antibodies.

Characteristics of Monoclonal Antibodies Against Human Chorionic Gonadotrophin

1982 ◽  
pp. 837-842 ◽  
Author(s):  
A.M.G. BOSCH ◽  
W. STEVENS ◽  
A. SCHUURS ◽  
O. SCHÖNHERR ◽  
H. ROELOFS
Keyword(s):  
Monoclonal Antibodies ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin

A COMPARISON BETWEEN CHORIONIC GONADOTROPHINS EXTRACTED FROM HUMAN, RHESUS MONKEY AND MARMOSET PLACENTAE

1972 ◽  
Vol 55 (2) ◽  
pp. 363-368 ◽  
Author(s):  
BRUCE HOBSON ◽  
LEIF WIDE
Keyword(s):  
Biological Activity ◽  
Rhesus Monkey ◽  
Human Placenta ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Antigenic Determinants

SUMMARY Evidence is provided to show that chorionic gonadotrophins extracted from the human, rhesus monkey and marmoset placentae have antigenic determinants in common. Similar slopes were obtained for these gonadotrophins in a radioimmunoassay for human chorionic gonadotrophin (HCG). The biological activity of the monkey gonadotrophins was neutralized by anti-HCG serum. When the gonadotrophic activity of the monkey placental extracts was assayed biologically and immunologically, using HCG as a standard, similar results were obtained. Higher values were obtained by the immunoassay than by the bioassay when extracts of human placenta were assayed using the same HCG standard.

scholarly journals Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies

2016 ◽  
Vol 82 (8) ◽  
pp. 2300-2311 ◽  
Author(s):  
Eva J. Scharinger ◽  
Richard Dietrich ◽  
Ina Kleinsteuber ◽  
Erwin Märtlbauer ◽  
Kristina Schauer
Keyword(s):  
Monoclonal Antibodies ◽  
Rapid Detection ◽  
Foodborne Pathogen ◽  
Cross Reactivity ◽  
Antigenic Determinants ◽  
Powdered Infant Formula ◽  
Bacterial Cells ◽  
Cronobacter Sakazakii ◽  
Content Type ◽  
Cell Counts

ABSTRACTCronobacter sakazakiiis a foodborne pathogen associated with rare but often lethal infections in neonates. Powdered infant formula (PIF) represents the most frequent source of infection. Out of the identified serotypes (O1 to O7), O1, O2, and O3 are often isolated from clinical and PIF samples. Serotype-specific monoclonal antibodies (MAbs) suitable for application in enzyme immunoassays (EIAs) for the rapid detection ofC. sakazakiihave not yet been developed. In this study, we created specific MAbs with the ability to bind toC. sakazakiiof serotypes O1, O2, and O3. Characterization by indirect EIAs, immunofluorescence, motility assays, and immunoblotting identified lipopolysaccharide (LPS) and exopolysaccharide (EPS) as the antigenic determinants of the MAbs. The established sandwich EIAs were highly sensitive and were able to detect between 2 × 103and 9 × 106CFU/ml. Inclusivity tests confirmed that 93% of serotype O1 strains, 100% of O2 strains, and 87% of O3 strains were detected at low cell counts. No cross-reactivity with >100 strains ofCronobacterspp. and otherEnterobacteriaceaewas observed, except for that withC. sakazakiiserotype O3 andCronobactermuytjensiiserotype O1. Moreover, the sandwich EIAs detectedC. sakazakiiin PIF samples artificially contaminated with 1 to 10 bacterial cells per 10 g of sample after 15 h of preenrichment. The use of these serotype-specific MAbs not only allows the reliable detection ofC. sakazakiistrains but also enables simultaneous serotyping in a simple sandwich EIA method.

PRODUCTION OF ANTISERA AGAINST HIGHLY PURIFIED HUMAN FOLLICLE-STIMULATING HORMONE, LUTEINIZING HORMONE AND THYROID-STIMULATING HORMONE

1976 ◽  
Vol 70 (3) ◽  
pp. 335-344 ◽  
Author(s):  
J. I. THORELL ◽  
B. HOLMSTRÖM
Keyword(s):  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Thyroid Stimulating Hormone ◽  
High Titre ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Antigenic Determinants ◽  
High Avidity ◽  
Cross Reactions ◽  
Stimulating Hormone

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.

scholarly journals Molar pregnancy and thyroid storm - literature review

2017 ◽  
Vol 23 (3) ◽  
pp. 121-125 ◽  
Author(s):  
G. A. Filipescu ◽  
Oana Alina Solomon ◽  
Nicoleta Clim ◽  
Amelia Milulescu ◽  
Andreea Gratiana Boiangiu ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Literature Review ◽  
Follicle Stimulating Hormone ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Thyroid Storm ◽  
Unusual Complication ◽  
Molar Pregnancy ◽  
Dilation And Curettage

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.

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