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Steroids ; 64(4): 259-65, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10399882

ABSTRACT

Four monoclonal antibodies to human sex hormone-binding globulin were raised and characterized. Three of the four antibodies recognised different antigenic determinants on SHBG. Two of the distinct antibodies were useful for Western blotting and recognized a major 48 kDa band in human plasma as well as a 46 kDa minor component. Carbohydrate residues do not form part of the antigenic determinants of these two antibodies, although one of these showed increased signal following removal of N-linked oligosaccharides. Some of the antibodies were selected to form a basis of a same-day, non-competitive, enzyme-linked immunosorbent assay (ELISA) for SHBG in plasma. The assay employs a purified IgG2a SHBG monoclonal antibody adsorbed to the wells of a microtitre plate. After blocking any further adsorption to the plate, standards or diluted patient samples were added for a 5-h incubation at room temperature, after which the plate was washed and antibody-bound SHBG was detected with an anti-SHBG IgG1 monoclonal antibody followed by peroxidase-labeled antimouse-IgG1 and o-phenylenediamine substrate. The assay correlated well with an existing 2-day ELISA for SHBG in plasma using polyclonal antibodies and also correlated with a dihydrosterone (DHT) ligand-binding assay. The monoclonal antibody-based ELISA shows excellent performance characteristics and is unaffected by added testosterone or estradiol.


Subject(s)
Antibodies, Monoclonal , Enzyme-Linked Immunosorbent Assay/methods , Sex Hormone-Binding Globulin/analysis , Adult , Animals , Antibody Specificity , Antigens/analysis , Antigens/immunology , Blotting, Western , Electrophoresis, Polyacrylamide Gel , Female , Humans , Hybridomas/immunology , Male , Mice , Pregnancy , Reference Values , Sex Hormone-Binding Globulin/immunology
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