Detection of Biased Agonist Activation
First Claim
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1. A method of determining if a ligand or stimulus is a biased agonist comprising:
- a) immobilizing or placing one or more cells on a surface of a colorimetric resonant reflectance optical biosensor;
b) applying a ligand or stimulus to the one or more cells;
c) detecting colorimetric resonant reflectance optical peak wavelength values (PWVs) for the one or more cells over time; and
d) comparing the PWVs of step (c) to PWVs over time of a full agonist acting on the same or similar one or more cells of step (a);
wherein, the ligand or stimulus is a biased agonist if the PWVs of the ligand or stimulus over time are smaller in magnitude or later in onset or both than the PWVs over time of the full agonist.
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Abstract
The invention provides single assay methods to detect the activity of biased agonists or agonists that are less than full agonists on cells.
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Citations
20 Claims
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1. A method of determining if a ligand or stimulus is a biased agonist comprising:
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a) immobilizing or placing one or more cells on a surface of a colorimetric resonant reflectance optical biosensor; b) applying a ligand or stimulus to the one or more cells; c) detecting colorimetric resonant reflectance optical peak wavelength values (PWVs) for the one or more cells over time; and d) comparing the PWVs of step (c) to PWVs over time of a full agonist acting on the same or similar one or more cells of step (a); wherein, the ligand or stimulus is a biased agonist if the PWVs of the ligand or stimulus over time are smaller in magnitude or later in onset or both than the PWVs over time of the full agonist. - View Dependent Claims (2, 3, 4, 5)
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6. A method of determining if a ligand or stimulus is a not a full agonist comprising:
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a) immobilizing or placing one or more cells on a surface of a colorimetric resonant reflectance optical biosensor; b) applying a ligand or stimulus to the one or more cells; c) detecting colorimetric resonant reflectance optical peak wavelength values (PWVs) for the one or more cells over time; and d) comparing the PWVs of step (c) to PWVs over time of a full agonist acting on the same or similar one or more cells of step (a); wherein, the ligand or stimulus is not a full agonist if the PWVs of the ligand or stimulus over time are smaller in magnitude or later in onset or both than the PWVs over time of the full agonist. - View Dependent Claims (7, 8, 9, 10)
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11. A method of determining if a ligand or stimulus is a biased agonist comprising:
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a) immobilizing or placing one or more cells on a surface of a grating based waveguide biosensor; b) applying a ligand or stimulus to the one or more cells; c) detecting changes in effective refractive indices for the one or more cells over time; and d) comparing the effective refractive indices of step (c) to effective refractive indices over time of a full agonist acting on the same or similar one or more cells of step (a); wherein, the ligand or stimulus is a biased agonist if the effective refractive indices of the ligand or stimulus over time are smaller in magnitude or later in onset or both than the effective refractive indices over time of the full agonist. - View Dependent Claims (12, 13, 14, 15)
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16. A method of determining if a ligand or stimulus is a not a full agonist comprising:
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a) immobilizing or placing one or more cells on a surface of a grating based waveguide biosensor; b) applying a ligand or stimulus to the one or more cells; c) detecting changes in effective refractive indices for the one or more cells over time; and d) comparing the effective refractive indices of step (c) to effective refractive indices over time of a full agonist acting on the same or similar one or more cells of step (a); wherein, the ligand or stimulus is not a full agonist if the effective refractive indices of the ligand or stimulus over time are smaller in magnitude or later in onset or both than the effective refractive indices over time of the full agonist. - View Dependent Claims (17, 18, 19, 20)
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Specification