New cell line stably expressing both beta-adrenergic receptor and a cyclic adenosine monophosphate sensor, useful for diagnosing beta-adrenergic receptor-mediated disease including e.g. infectious and non-infectious heart diseases

摘要

Cell line stably expressing both beta -adrenergic receptor and a cyclic adenosine monophosphate (cAMP)-sensor, is new. Independent claims are included for: (1) determining the concentration of cAMP using the stable cell line, comprising measuring/detecting fluorescence resonance energy transfer- or bioluminescence resonance energy transfer-signals and determining the cAMP concentration by comparing the values of the fluorescence emission with a standard curve; (2) identifying molecules or substances that bind to beta -adrenergic receptors, comprising contacting the cell line with molecules or substances to be tested and measuring a change of light intensity, which is emitted by the detectable marker that comprises cAMP sensor as a chimeric peptide; (3) screening molecules or substances that are activators (agonists) or inhibitors (antagonists) of a beta -adrenergic receptor, comprising contacting the cell line with molecules or substances to be tested, measuring and/or detecting a change in light intensity that is emitted by the detectable marker comprising cAMP sensor as chimeric peptide, and comparing the change in light intensity with a standard curve, as measured in the absence of the candidate molecules/substances; (4) diagnosing beta -adrenergic receptor-mediated disease, comprising providing a sample from a patient, contacting the cell line with molecules or substances to be tested, measuring and/or detecting a change in light intensity emitted by the detectable marker comprising cAMP sensor as a chimeric peptide and comparing the change in light intensity with a standard curve, as measured in the absence of the candidate molecules/compounds, where an elevated content of cAMP in cells of stably transformed cell line as reduction on sample material is compared to the standard curve indicating that the patient suffering from beta -adrenergic receptor-mediated disease; (5) progression of beta -adrenergic receptor-mediated disease for a period of at least 3 months using the stable cell line, comprising providing a sample from a patient, contacting the cell line with molecules or substances to be tested from the sample material, measuring and/or detecting a change in the light intensity emitted by the detectable marker comprising cAMP-sensor as a chimeric peptide and comparing the change in light intensity with a standard curve, as measured in the absence of the candidate molecules/substances, where an elevated content of cAMP in cells of stably transformed cell line, as reduction on the sample material is compared to the standard curve indicating that the patient suffering from adrenergic receptor-mediated disease, and the content of cAMP is longitudinally measured; (6) evaluating a selected dose of beta -adrenergic receptor homologs in the treatment of beta -adrenergic receptor-mediated disease using the stable cell line, comprising providing a sample from a patient, contacting the cell line with molecules or substances to be tested from the sample, measuring and/or detecting a change in light intensity that is emitted by the detectable marker comprising cAMP sensor as chimeric peptide and comparing the change in light intensity with a standard curve, as measured in the absence of the candidate molecules/substances, where an elevated content of cAMP in the cells of stably transformed cell line as reduction to the sample material is compared to the standard curve indicating that the selected dose for treatment of beta -adrenergic receptor-mediated disease is not sufficient, and the dose is selected so that no increased content of cAMP in the cells of the cell line is detected compared to the standard curve; (7) a diagnostic composition comprising the stable cell line and activator (agonist); and (8) a kit comprising the stable cell line.