Anti-equine Fc epsilon receptor alpha chain antibodies and methods to detect IgE
View Patent ↗The present invention relates to equine Fc epsilon receptor alpha chain nucleic acid molecules, proteins encoded by such nucleic acid molecules, antibodies raised against such proteins, and inhibitors of such proteins. The present invention also includes methods to detect IgE using such proteins and antibodies. Also included in the present invention are therapeutic compositions comprising such proteins, nucleic acid molecules, antibodies and/or inhibitory compounds as well as the use of such therapeutic compositions to mediate Fc epsilon receptor-mediated biological responses.
1. A method to detect IgE comprising: (a) contacting an isolated equine Fc ε Rα molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12 and amino acid sequences at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, and SEQ ID NO:12, wherein said molecule comprises an IgE binding domain and binds to IgE, with a putative IgE-containing composition under conditions suitable for formation of a Fc ε Rα molecule:IgE complex; and
(b) determining the presence of IgE by detecting said Fc ε Rα molecule:IgE complex, the presence of said Fc ε Rα molecule:IgE complex indicating the presence of IgE.
2. The method of claim 1 , wherein said Fc ε Rα molecule comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, and SEQ ID NO:12.
3. The method of claim 1 , wherein said Fc ε Rα molecule is encoded by a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO: 11.
4. The method of claim 1 , wherein said Fc ε Rα molecule is conjugated to a detectable marker.
5. The method of claim 1 , wherein said Fc ε Rα molecule is conjugated to a detectable marker selected from the group consisting of a radioactive label, an enzyme, a fluorescent label, a chemiluminescent label, a chromophoric label and ligand.
6. The method of claim 1 , wherein said Fc ε Rα molecule is conjugated to a detectable marker selected from the group consisting of fluorescein, a radioisotope, a phosphatase, biotin, biotin-related compounds, avidin, avidin-related compounds and a peroxidase.
7. The method of claim 4 , wherein said detectable marker is connected to said Fc ε Rα molecule by chemical conjugation or recombinant DNA technology.
8. The method of claim 1 , wherein said Fc ε Rα molecule is conjugated to biotin.
9. The method of claim 1 , wherein said putative IgE-containing composition comprises a composition selected from the group consisting of blood, serum, plasma, urine, tears, aqueous humor, cerebrospinal fluid, saliva, lymph, nasal secretions, tracheobronchial aspirates, milk and feces.
10. The method of claim 1 , wherein said putative IgE-containing composition comprises serum.
11. The method of claim 1 , wherein said putative IgE-containing composition comprises a cell that produces IgE.
12. The method of claim 1 , further comprising the step selected from the group consisting of immobilizing said Fc ε Rα molecule on a substrate prior to performing step (a) to form a Fc ε Rα molecule-immobilized substrate; and binding said putative IgE-containing composition on a substrate prior to performing step (a) to form a putative IgE-containing composition-bound substrate, wherein said substrate is selected from the group consisting of a non-coated substrate, a Fc ε Rα molecule-immobilized substrate, an antigen-immobilized substrate and an anti-IgE antibody-immobilized substrate.
13. The method of claim 12 , wherein said substrate comprises a material selected from the group consisting of plastic, glass, gel, celluloid, paper and particulate material.
14. The method of claim 12 , wherein said substrate is selected from the group consisting of a well, a plate, a dipstick, a bead, a lateral flow apparatus, a membrane, a filter, a tube, a dish, a celluloid-type matrix and a magnetic particle.
15. The method of claim 1 , wherein said step of detecting comprises performing assays selected from the group consisting of enzyme-linked immunoassay, radioimmunoassay, immunoprecipitation, fluorescence immunoassay, chemiluminescent assay, immunoblot assay, lateral flow assay, agglutination assay and particulate-based assay.
16. The method of claim 1 , wherein said method is performed in solution.