Methods to identify compounds that modulate RAGE
View Patent ↗Provided are methods to detect of modulators of the Receptor for Advanced Glycated Endproducts (RAGE). The invention comprises a method for detection of RAGE modulators comprising: adsorbing a RAGE ligand onto a solid surface; adding a compound of interest and a protein comprising RAGE or fragment thereof, to the preadsorbed ligand; adding an antibody which binds to RAGE or fragment thereof and a secondary antibody which binds to the anti-RAGE antibody; measuring the secondary antibody bound to the anti-RAGE antibody; and comparing the amount of RAGE bound to the ligand in the presence of varying amounts of the compound of interest. In an embodiment, the fragment of RAGE is sRAGE. In one aspect, the invention use of compounds detected by the method for treatment of AGE-related syndromes including complications associated with diabetes, kidney failure, lupus nephritis or inflammatory lupus nephritis, amyloidoses, Alzheimer's disease, cancer, inflammation, and erectile dysfunction.
1. A method for detection of RAGE ligands comprising:
adsorbing a RAGE ligand onto a solid surface;
adding a compound of interest and a protein comprising sRAGE as defined by the amino acid sequence SEQ ID NO: 2, to the preadsorbed ligand;
adding anti-RAGE antibody which binds to the sRAGE protein;
determining the amount of sRAGE protein bound to the ligand by measuring the amount of anti-RAGE antibody bound; and
comparing the amount of sRAGE protein bound to the ligand in the presence of varying amounts of the compound of interest.
2. The method of claim 1 , wherein the anti-RAGE antibody comprises a monoclonal antibody.
3. The method of claim 1 , herein the anti-RAGE antibody comprises a polyclonal antibody.
4. The method of claim 1 , further comprising adding a second antibody which recognizes the anti-RAGE antibody.
5. The method of claim 4 , wherein the anti-RAGE antibody and secondary antibody are allowed to complex prior to being added.
6. The method of claim 4 , further comprising performing a calorimetric assay for the secondary antibody.
7. The method of claim 1 , wherein the solid surface comprises a microtiter well.
8. The method of claim 1 , wherein the solid surface comprises a dip-stick.
9. The method of claim 1 , wherein the ligand comprises S100b.
10. The method of claim 1 , wherein the compound of interest comprises a synthetic peptide.
11. The method of claim 1 , wherein the compound of interest comprises a peptidomimetic.
12. The method of claim 1 , wherein the compound of interest comprises an organic compound.
13. The method of claim 1 , wherein the compound of interest comprises an inorganic compound.
14. The method of claim 1 , wherein the compound of interest comprises a lipid.
15. The method of claim 1 , wherein the compound of interest comprises a carbohydrate.
16. The method of claim 1 , wherein compound of interest comprises a nucleic acid.
17. A method for detection of RAGE ligands comprising:
adsorbing a RAGE ligand onto a solid surface;
adding a compound of interest and a protein comprising RAGE as defined by the amino acid sequence SEQ ID NO: 2 to the preabsorbed ligand;
adding anti-RAGE antibody which binds to the sRAGE protein and a secondary antibody which binds to the anti-RAGE antibody;
measuring the secondary antibody bound to the anti-RAGE antibody; and
comparing the amount of sRAGE protein bound to the ligand in the presence of varying amounts of the compound of interest.
18. The method of claim 17 , further comprising performing a calorimetric assay for the secondary antibody.
19. The method of claim 17 , wherein the anti-RAGE antibody and secondary antibody are allowed to complex prior to being added to the reaction.
20. The method of claims 17 , wherein the solid surface comprises a reaction vessel.
21. The method of claim 17 , wherein the solid surface comprises a dip-stick.