Chemistry: analytical and immunological testing – Biospecific ligand binding assay
Patent
1987-10-20
1991-12-10
Schwartz, Richard A.
Chemistry: analytical and immunological testing
Biospecific ligand binding assay
435 697, 4351723, C12Q 100, C12Q 125
Patent
active
050717731
ABSTRACT:
The present invention discloses two hormone receptor-related bioassays. The first bioassay is useful for determining whether a protein suspected of being a hormone receptor has transcription-activating properties of a hormone receptor. The second bioassay is useful for evaluating whether compounds are functional ligands for receptor proteins. According to the first bioassay, cells that contain non-endogenous DNA which expresses a protein suspected of being a hormone receptor and which contain a DNA sequence encoding an operative hormone responsive promoter/enhancer element linked to an operative reporter gene, are cultured, the culturing being conducted in a culture medium containing a known hormone, or an analog thereof. The cultured cells are then monitored for induction of the product of the reporter gene as an indication of functional transcription-activating binding between the hormone or hormone analog and the protein suspected of being a hormone receptor. According to the second bioassay, cells that contain non-endogenous DNA which expresses hormone receptor or a functional engineered or modified form thereof, and which also contain a DNA sequence encoding an operative hormone responsive promoter/enhancer element linked to an operative reporter gene, are cultured, the culturing being conducted in culture medium containing at least one compound whose ability to functionally bind the receptor protein is sought to be determined. The cultured cells are then monitored for induction of the product of the report gene as an indicator of functional binding between the compound and the receptor.
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Arriza Jeffrey
Evans Ronald M.
Giguere Vincent
Hollenberg Stanley M.
Ong Estelita S.
Schwartz Richard A.
The Salk Institute for Biological Studies
Ulm John D.
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