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• W1996976402 abstract "To probe possible effects of carbohydrate chains in the conformation of pituitary glycoprotein hormones, two radiolabeled derivatives of human thyroid-stimulating hormone (hTSH), either partially deglycosylated in the β-subunit or fully deglycosylated in both the α- and β-subunits, were compared to the native hormone for binding to monoclonal as well as polyclonal antibodies. Monoclonal antibodies were screened for their ability to bind the intact hormone (anti-hTSH), hTSH and its freeα-subunit (anti-α) or its free β-subunit (anti-β). A panel of 14 monoclonal antibodies directed against at least eight out of the 12 epitopes known to be present in the hormone was tested in solid-phase assays for their capacity to bind intact and deglycosylated forms of hTSH. All of them displayed identical recognition of native and partially deglycosylated 125 I-hTSH. In contrast, binding of fully deglycosylated 125I-hTSH to anti-hTSH and anti-β antibodies was dramatically lost while that of anti-α was preserved. This clearly indicates that most of the epitopes specific for subunit association as well as those present on the β-subunit are glycosylation dependent. No alteration was found in antibody recognition following deglycosylation of free individual subunits, indicating that the carbohydrate effect can only occur in the combined dimer. Using polyclonal antisera raised against the International Reference Preparations, we found that the deglycosylated hormone could be bound by the anti-β antiserum although at a much lower dilution than the native antigen, suggesting the presence of at least one glycosylation-independent epitope in the β-subunit. Competitive binding assays revealed that deglycosylated hTSH is 5 times less immunoreactive toward the anti-β compared to the anti-α antiserum. The current data thus demonstrate the presence of the glycosylation-independent epitopes in the α-subunit of hTSH and the localization of most of the glycosylation-dependent domains in the β-subunit." @default.
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• W1996976402 date "1990-10-01" @default.
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• W1996976402 title "Carbohydrate-dependent epitope mapping of human thyrotropin" @default.
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• W1996976402 doi "https://doi.org/10.1016/0303-7207(90)90040-f" @default.
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