Chimeric molecules and novel assay system
OwnerNeil Stahi, Margaret Karow +1inventors
InventorsNeil Stahi, Margaret Karow, George D. Yancopoulos
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Abstract
The present invention provides for a method of identifying a ligand for a receptor comprising contacting a cell expressing a cell surface FcεR1 molecule with a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE constant or Fc region, under conditions whereby the chimeric polypeptide molecule binds to the FcεR1 molecule to form a complex; contacting the cell bearing the complex with a ligand; and detecting or measuring ligand binding to the complex. The invention further provides for chimeric polypeptide molecules, the nucleic acids encoding the chimeric polypeptide molecules, and cell lines expressing the chimeric polypeptide molecules.
Claims54
A nucleic acid encoding a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE constant region.
A nucleic acid encoding a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE Fc region.
A nucleic acid encoding a chimeric polypeptide molecule comprising a protein interacting region and an IgE constant region.
A nucleic acid encoding a chimeric polypeptide molecule comprising a protein interacting region and an IgE Fo region.
A chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE constant region.
A chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE Fc region.
A chimeric polypeptide molecule comprising a protein interacting region and an IgE constant region.
A chimeric polypeptide molecule comprising a protein interacting region and an IgE Fc region.
An assay system comprising
- a) a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor fused to an IgE constant region;
- and
- b) a means of detecting or measuring ligand binding to the chimeric polypeptide molecule.
An assay system comprising
- a) a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor fused to an IgE Fc region;
- and
- b) a means of detecting or measuring ligand binding to the chimeric polypeptide molecule.
An assay system comprising
- a) a chimeric polypeptide molecule comprising a protein interacting region fused to an IgE constant region;
- and
- b) a means of detecting or measuring ligand binding to the chimeric polypeptide molecule.
An assay system comprising
- a) a chimeric polypeptide molecule comprising a protein interacting region fused to an IgE Fc region;
- and
- b) a means of detecting or measuring ligand binding to the chimeric polypeptide molecule.
A cell line stably expressing a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE constant region.
A cell line stably expressing a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE Fc region.
A cell line stably expressing a chimeric polypeptide molecule comprising a protein interacting region and an IgE constant region.
A cell line stably expressing a chimeric polypeptide molecule comprising a protein interacting region and an IgE Fc region.
A method of identifying a ligand for a receptor comprising:
- (a) contacting a cell expressing a cell surface FcεR1 molecule with a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE constant region, under conditions whereby said chimeric polypeptide molecule binds to said FcεR1 molecule to form a complex;
- (b) contacting said cell bearing said complex with a ligand;
- and
- (c) detecting or measuring said ligand binding to said complex.
A method of identifying a ligand for a receptor comprising:
- (a) contacting a cell expressing a cell surface FcεR1 molecule with a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and an IgE Fc region, under conditions whereby said chimeric polypeptide molecule binds to said FcεR1 molecule to form a complex;
- (b) contacting said cell bearing said complex with a ligand;
- and
- (c) detecting or measuring said ligand binding to said complex.
A method of identifying a candidate antagonist to a receptor comprising:
- (a) contacting a cell expressing a cell surface FcεR1 molecule with:
- (i) a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and the IgE constant region;
- and
- (ii) a ligand for said receptor, in the presence and absence of a candidate antagonist for said receptor, and under conditions whereby said chimeric polypeptide molecule binds to said cell surface FcεR1 molecule to provide a cell bearing complexes of said FcεR1 molecule and said chimeric polypeptide molecule;
- and
- (b) measuring binding of said ligand to said complex in the presence and absence of said candidate antagonist, wherein decreased ligand binding in the presence of said candidate antagonist is indicative of identification of an antagonist.
A method of identifying a candidate antagonist to a receptor comprising:
- (a) contacting a cell expressing a cell surface FcεR1 molecule with:
- (i) a chimeric polypeptide molecule comprising an extracellular ligand binding domain of a receptor and the IgE Fc region;
- and
- (ii) a ligand for said receptor, in the presence and absence of a candidate antagonist for said receptor, and under conditions whereby said chimeric polypeptide molecule binds to said cell surface FcεR1 molecule to provide a cell bearing complexes of said FcεR1 molecule and said chimeric polypeptide molecule;
- and
- (b) measuring binding of said ligand to said complex in the presence and absence of said candidate antagonist, wherein decreased ligand binding in the presence of said candidate antagonist is indicative of identification of an antagonist.
Dependencies are inferred from the claim text.