IL-17F-specific capture agents, compositions, and methods of using and making
The present application provides stable peptide-based IL-17F capture agents and methods of use as detection agents. The application further provides methods of manufacturing IL-17F capture agents.
1. A stable, synthetic capture agent that specifically binds IL-17F, wherein the capture agent comprises a first ligand having binding affinity for a first epitope on an IL-17F, a second ligand having binding affinity for a second epitope on the IL-17F, and a linker covalently connecting the first ligand to the second ligand,
wherein the first ligand specifically binds to the first epitope and the second ligand specifically binds to the second epitope, wherein the first epitope comprises the amino acid sequence FFQKPES (SEQ ID NO:1), wherein the first ligand comprises the amino acid sequence FYKTH (SEQ ID NO:5), FYKQH (SEQ ID NO:6), FYLTH (SEQ ID NO:7), FYLQH (SEQ ID NO:8), RRATS (SEQ ID NO:9), or RRAQS (SEQ ID NO:10),
wherein the second epitope comprises the amino acid sequence NENQRVS (SEQ ID NO:3), wherein the second ligand comprises the amino acid sequence KYGEV (SEQ ID NO:11), LYGEV (SEQ ID NO:12), VHKSG (SEQ ID NO:13), VHLSG (SEQ ID NO:14), QKHGP (SEQ ID NO:15), TKHGP (SEQ ID NO:16), QLHGP (SEQ ID NO:17), TLHGP (SEQ ID NO:18), YDLQR (SEQ ID NO:19), YDLTR (SEQ ID NO:20), YDKQR (SEQ ID NO:21), YDKTR (SEQ ID NO:22), KKGWP (SEQ ID NO:23), KLGWP (SEQ ID NO:24), LKGWP (SEQ ID NO:25), LLGWP (SEQ ID NO:26), RSYNL (SEQ ID NO:27), and RSYNK (SEQ ID NO:28).
2. The capture agent of claim 1 , wherein the capture agent is selective for IL-17F over IL-17A.
3. The capture agent of claim 1 , wherein the first epitope comprises the amino acid sequence FFQKPESCPPVPGG (SEQ ID NO:2).
4. The capture agent of claim 1 , wherein the second epitope comprises the amino acid sequence GIINENQRVS (SEQ ID NO:4).
5. The capture agent of claim 1 , wherein the first ligand comprises the amino acid sequence RRATS (SEQ ID NO:9) or RRAQS (SEQ ID NO:10).
6. The capture agent of claim 1 , wherein the first ligand comprises the sequence RRATS (SEQ ID NO:9).
7. The capture agent of claim 1 , wherein the first ligand comprises the sequence RRAQS (SEQ ID NO:10).
8. The capture agent of claim 1 , wherein the first ligand is cyclic.
9. The capture agent of claim 1 , wherein the first ligand comprises a 1,4-substituted-1,2,3-triazole residue (Tz4) or a 1,5-substituted-1,2,3-triazole residue (Tz5).
10. The capture agent of claim 9 , wherein the triazole residue is a 1,4-substituted-1,2,3-triazole (Tz4) residue.
11. The capture agent of claim 1 , wherein the second ligand comprises the amino acid sequence TKHGP (SEQ ID NO:16), QKHGP (SEQ ID NO:15), KKGWP (SEQ ID NO:23), or RSYNK (SEQ ID NO:28).
12. The capture agent of claim 1 , wherein the second ligand comprises the amino acid sequence TKHGP (SEQ ID NO:16).
13. The capture agent of claim 1 , wherein the second ligand comprises the amino acid sequence QKHGP (SEQ ID NO:15).
14. The capture agent of claim 1 , wherein the second ligand comprises the amino acid sequence KKGWP (SEQ ID NO:23).
15. The capture agent of claim 1 , wherein the second ligand comprises the amino acid sequence RSYNK (SEQ ID NO:28).
16. The capture agent of claim 1 , wherein the second ligand is cyclic.
17. The capture agent of claim 1 , wherein the second ligand comprises a 1,4-substituted-1,2,3-triazole residue (Tz4) or a 1,5-substituted-1,2,3-triazole residue (Tz5).
18. The capture agent of claim 17 , wherein the triazole residue is a 1,4-substituted-1,2,3-triazole residue (Tz4).
19. The capture agent of claim 1 , wherein the linker is divalent.
20. The capture agent of claim 1 , wherein the length of the linker corresponds to distance between the first epitope and the second epitope.
21. The capture agent of claim 20 , wherein the length of the linker is from ˜4.4 Å to ˜26.4 Å, from ˜8.8 Å to ˜26.4 Å or from ˜7 Å to ˜15 Å.
22. The capture agent of claim 20 , wherein the length of the linker is ˜15 Å.
23. The capture agent of claim 1 , wherein the linker comprises one or more repeat units of ethylene glycol.
24. The capture agent of claim 1 , wherein the linker comprises a peptide.
25. The capture agent of claim 1 , wherein the first ligand comprises the sequence RRATS (SEQ ID NO:9) and the second ligand comprises the sequence QKHGP (SEQ ID NO:15).
26. The capture agent of claim 1 , wherein the first ligand comprises the sequence RRATS (SEQ ID NO:9) and the second ligand comprises the sequence RSYNK (SEQ ID NO:28).
27. The capture agent of claim 26 , wherein the first and second ligands are cyclic and comprise a Tz4 residue.
28. The capture agent of claim 27 , wherein the linker is glycine.
29. The capture agent of claim 27 , wherein the linker is PEG 1 .
30. The capture agent of claim 27 , wherein the linker is PEG 2 .
31. The capture agent of claim 27 , wherein the linker is PEG 3 .
32. The capture agent of claim 27 , wherein the linker is PEG 4 .
33. The capture agent of claim 27 , wherein the linker is PEG 5 .
34. The capture agent of claim 1 , wherein the capture agent is labeled with a detectable moiety.
35. The capture agent of claim 34 , wherein the detectable moiety is selected from the group consisting of biotin, copper-DOTA, biotin-PEG3, aminooxyacetate, 19 FB, 18 FB and FITC-PEG 3 .
36. The capture agent of claim 34 , wherein the detectable moiety is selected from the group consisting of 64 Cu DOTA, 68 Ga DOTA, 68 Ga NOTA, 18 F, Al 18 F NOTA, 64 Cu, 68 Ga, 89 Zr, 124 I, 86 Y, 94m Tc, 110m In, 11 C and 76 Br.
37. A stable, synthetic capture agent that specifically binds IL-17F having a structure selected from the group consisting of:
38. A stable, synthetic capture agent that specifically binds IL-17F having a structure selected from the group consisting of:
39. A method of detecting IL-17F in a biological sample, comprising the steps of contacting the biological sample with one or more of the capture agents of claim 1 under conditions allowing binding of the capture agents to IL-17F present in the sample to form a complex, and detecting the complex to detect IL-17F in the biological sample.
40. The capture agent of claim 1 , wherein the length of the linker corresponds to distance between the first epitope and the second epitope, wherein the linker comprises one or more repeat units of ethylene glycol, a peptide, or both.
41. The capture agent of claim 40 , wherein the length of the linker is from ˜4.4 Å to ˜26.4 Å, from ˜8.8 Å to ˜26.4 Å or from ˜7 Å to ˜15 Å.
42. The capture agent of claim 40 , wherein the length of the linker is ˜15 Å.
43. The capture agent of claim 40 , wherein the linker is glycine.
44. The capture agent of claim 40 , wherein the linker is PEG 1 .
45. The capture agent of claim 40 , wherein the linker is PEG 2 .
46. The capture agent of claim 40 , wherein the linker is PEG 3 .
47. The capture agent of claim 40 , wherein the linker is PEG 4 .
48. The capture agent of claim 40 , wherein the linker is PEG 5 .
49. The capture agent of claim 40 , wherein the first and second ligands are cyclic and comprise a 1,4-substituted-1,2,3-triazole residue (Tz4) or a 1,5-substituted-1,2,3-triazole residue (Tz5).
50. A stable, synthetic capture agent that specifically binds IL-17F, wherein the capture agent comprises a first ligand having binding affinity for a first epitope on an IL-17F, a second ligand having binding affinity for a second epitope on the IL-17F, and a linker covalently connecting the first ligand to the second ligand,
wherein the first ligand comprises the sequence RRATS (SEQ ID NO:9) and the second ligand comprises the sequence RSYNK (SEQ ID NO:28), wherein the first and second ligands are cyclic and comprise a Tz4 residue, wherein the linker is glycine, PEG 1 , PEG 2 , PEG 3 , PEG 4 , or PEG 5 .