Method for multiple chemokine receptor screening for antagonists using RAM assay
First Claim
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1. A method for identifying a chemoattractant receptor antagonist, comprising:
- incubating a cell population comprising first and second chemoattractant receptors;
contacting the cell population with an inhibitory concentration of a ligand for the first chemoattractant receptor;
contacting the cell population with an inhibitory concentration of a ligand for the second chemoattractant receptor;
contacting the cell population with a candidate antagonist;
assaying migration of the cell population, wherein migration identifies the candidate antagonist as an antagonist of at least one of the first and second chemoattractant receptors; and
determining whether an identified antagonist is an antagonist for one of the first chemoattractant receptors, the second chemoattractant receptor, or both.
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Abstract
The present invention is directed to a modified cell migration assay allowing for improved identification and discrimination of chemokine receptor antagonists from non-specific migration blockers.
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Citations
60 Claims
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1. A method for identifying a chemoattractant receptor antagonist, comprising:
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incubating a cell population comprising first and second chemoattractant receptors;
contacting the cell population with an inhibitory concentration of a ligand for the first chemoattractant receptor;
contacting the cell population with an inhibitory concentration of a ligand for the second chemoattractant receptor;
contacting the cell population with a candidate antagonist;
assaying migration of the cell population, wherein migration identifies the candidate antagonist as an antagonist of at least one of the first and second chemoattractant receptors; and
determining whether an identified antagonist is an antagonist for one of the first chemoattractant receptors, the second chemoattractant receptor, or both. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. A method for identifying a chemoattractant receptor antagonist, comprising:
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incubating a first cell population and a second cell population, wherein the first cell population comprises a first chemoattractant receptor and wherein the second cell population comprises a second chemoattractant receptor;
contacting the first and the second cell populations with an inhibitory concentration of a ligand for the first chemoattractant receptor;
contacting the first and the second cell populations with an inhibitory concentration of a ligand for the second chemoattractant receptor;
contacting the first and the second cell populations with a candidate antagonist;
assaying migration of the first and the second cell populations, wherein migration identifies the candidate antagonist as an antagonist of at least one of the first and second chemoattractant receptors; and
determining whether an identified antagonist is an antagonist for one of the first chemoattractant receptors, the second chemoattractant receptor, or both. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54)
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55. A kit comprising
a cell migration apparatus, and at least one chemokine.
Specification