Methods of using chimeric receptors to identify autoimmune disease
View Patent ↗The present invention provides methods and compositions useful in the diagnosis and management of autoimmune diseases. In particular, the present invention provides improved methods and compositions for the diagnosis and management of Graves' disease. The methods of the present invention not only avoids the need for radioactivity and are much simpler, economical, and rapid than methods traditionally used for the diagnosis of Graves' disease, but also improve upon the sensitivity and detection abilities of previous luciferase-based autoantibody detection assays.
1. A recombinant chimeric thyroid stimulating hormone (TSH) receptor polypeptide encoded by nucleotides 45-2234 of SEQ ID NO:3.
2. A recombinant polypeptide consisting of a chimeric thyroid stimulating hormone (TSH) receptor, wherein said chimeric TSH receptor consists of:
an N-terminal portion of a human thyroid stimulating hormone receptor (hTSHR), wherein the N-terminal portion of the hTSHR consists of the amino acid sequence encoded by nucleotides 45-827 of SEQ NO:3;
a portion of a rat luteinizing hormone chorionic gonadotropin receptor (rLH/CGR), wherein the portion of the rLH/CGR consists of the amino acid sequence encoded by nucleotides 828 to 1046 of SEQ ID NO:3; and
a C-terminal portion of said hTSHR, wherein the C-terminal portion of the hTSHR consists of the amino acid sequence encoded by nucleotides 1047-2234 of SEQ ID NO:3.
3. A recombinant nucleic acid sequence which encodes said recombinant chimeric thyroid stimulating hormone (TSH) receptor polypeptide of claim 2 .
4. A nucleotide vector comprising a recombinant nucleic acid sequence which encodes said recombinant chimeric thyroid stimulating hormone (TSH) receptor polypeptide of claim 2 .
5. The vector of claim 4 , further comprising a promoter operably linked to the recombinant nucleic acid sequence.
6. The vector of claim 5 , wherein the promoter comprises a glycoprotein alpha subunit promoter.
7. The vector of claim 4 , further comprising a luciferase nucleic acid sequence.
8. The vector of claim 7 , wherein the luciferase nucleic acid sequence is derived from a firefly.
9. A cell line comprising a transfected vector, wherein the vector comprises the nucleic acid sequence of claim 3 .
10. The cell line of claim 9 , wherein the cell line is stably transfected with the vector.
11. The cell line of claim 9 , wherein the cell line is selected from the group consisting of CHO cells and RD cells.
12. A kit comprising the nucleic acid sequence of claim 3 .
13. The kit of claim 12 , further comprising a cell line which is transfected with the nucleic acid sequence.
14. The kit of claim 13 , further comprising polyethylene glycol.
15. The kit of claim 12 , wherein the nucleic acid sequence is integrated into a vector.
16. The kit of claim 15 , wherein the vector further comprises a luciferase gene and a promoter wherein the promoter is operably linked to the nucleic acid sequence.