Antigenic features of human follicle stimulating hormone delineated by monoclonal antibodies and construction of an immunoradiomometric assay
The characterization of human (h) FSH with 181 monoclonal antibodies (MCA) allowed the elucidation of its antigenic topography. One- and two-site, limited as well as excess reagent type radioimmuno- and enzymoimmunoassays revealed three main categories of MCA molecular binding specificities; two thirds of all antibodies were directed against the alpha-subunit and one fourth toward the beta-chain, and less than one tenth recognized the conformationally (c) intact holohormone. With high frequency immunization schedules these specificities were shifted toward a higher proportion of beta-MCA. On the basis of intra- and interspecies cross-reaction studies as well as epitope contiguity analyses by sandwich assays, the three main categories could be further subdivided into nine epitopes: 1) five epitopes associated with the alpha-subunit, two of which were suprisingly shared by other species, and two being iodination sensitive, 2) two evolutionary conserved structures on the beta-subunit, adjacent to each other, and 3) two c-determinants, one of these present also on hTSH. The epitopes were arranged in three major antigenic domains, which seems to be a common homologous construction principle of the four human glycoprotein hormones: a central domain, consisting of three identically arranged alpha- and similarly located c-epitopes, is flanked by a single spatially distinct domain on each subunit. The establishment of an epitope map was followed by the construction of an immunoradiometric assay with a sensitivity of 0.25 ng hFSH/ml and an apparent cross-reactivity vs. hLH, hTSH, and hCG of less than 1%.
- Research Organization:
- Immunoendocrinology Research Unit, Innsbruck (Austria)
- OSTI ID:
- 6545038
- Journal Information:
- Endocrinology; (United States), Vol. 123:5
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
FSH
CHEMICAL COMPOSITION
MONOCLONAL ANTIBODIES
SPECIFICITY
ANTIGENS
CPB
ENZYME IMMUNOASSAY
HYBRIDOMAS
MICE
RADIOIMMUNOASSAY
ANIMAL CELLS
ANIMALS
ANTIBODIES
GONADOTROPINS
HORMONES
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
MAMMALS
PEPTIDE HORMONES
PITUITARY HORMONES
RADIOASSAY
RADIOIMMUNOLOGY
RODENTS
TRACER TECHNIQUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques