IL-17F and IL-17A-specific capture agents, compositions, and methods of using and making
The present application provides stable peptide-based IL-17F and IL-17A capture agents and methods of use as detection agents. The application further provides methods of manufacturing IL-17F capture agents.
1. A capture agent for a target, the capture agent comprising two or more ligands covalently linked to each other, wherein the ligands each specifically bind to one of two or more distinct epitopes of the target that are in different locations on the target,
wherein the capture agent comprises a first of the ligands having specific affinity for a first of the epitopes, a second of the ligands having specific affinity for a second of the epitopes, and a linker covalently connecting the first ligand to the second ligand,
wherein the target is IL-17A, IL-17F, or both IL-17A and IL-17F,
wherein the first epitope comprises the amino acid sequence FFQKPES (SEQ ID NO:1) or the amino acid sequence PNSEDKNFPRTVMVNL (SEQ ID NO:43), wherein the second epitope comprises the amino acid sequence NENQRVS (SEQ ID NO:3) or the amino acid sequence PNSEDKNFPRTVMVNL (SEQ ID NO:43),
wherein the first ligand comprises a first peptide having an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), rkhyh (SEQ ID NO:46), rrATS (SEQ ID NO:47), rrAQS (SEQ ID NO:48), rrats (SEQ ID NO:49), and rraqs (SEQ ID NO:50),
wherein the second ligand comprises a second peptide having an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of 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), RSYNK (SEQ ID NO:28), kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64), vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), rsynk (SEQ ID NO:76), rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46),
wherein the first ligand is cyclic, and wherein the second ligand is cyclic.
2. The capture agent of claim 1 , wherein the capture agent specifically binds IL-17F.
3. The capture agent of claim 1 , wherein the capture agent is selective for IL-17F over IL-17A.
4. The capture agent of claim 1 , wherein the first epitope comprises the amino acid sequence FFQKPESCPPVPGG (SEQ ID NO:2).
5. The capture agent of claim 1 , wherein the second epitope comprises the amino acid sequence GIINENQRVS (SEQ ID NO:4).
6. The capture agent of claim 1 , wherein the first ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rrATS (SEQ ID NO:47), rrAQS (SEQ ID NO:48), rrats (SEQ ID NO:49), and rraqs (SEQ ID NO:50).
7. The capture agent of claim 1 , wherein the second ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64)vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), and rsynk (SEQ ID NO:76).
8. The capture agent of claim 1 , wherein the first ligand comprises the sequence rrats (SEQ ID NO:49) and the second ligand comprises the sequence rsynk (SEQ ID NO:76).
9. The capture agent of claim 1 , having a structure selected from the group consisting of:
10. 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).
11. The capture agent of claim 10 , wherein the first ligand comprises a 1,4-substituted-1,2,3-triazole residue (Tz4).
12. The capture agent of claim 10 , wherein the first ligand comprises a 1,5-substituted-1,2,3-triazole residue (Tz5).
13. 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).
14. The capture agent of claim 13 , wherein the second ligand comprises a 1,4-substituted-1,2,3-triazole residue (Tz4).
15. The capture agent of claim 13 , wherein the second ligand comprises a 1,5-substituted-1,2,3-triazole residue (Tz5).
16. The capture agent of claim 1 , wherein the capture agent is labeled with a detectable moiety.
17. The capture agent of claim 16 , wherein the detectable moiety is selected from the group consisting of biotin, copper-DOTA, biotin-PEG3, aminooxyacetate, 19 FB, 18 FB and FITC-PEG3.
18. The capture agent of claim 16 , 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.
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 1 , wherein the length of the linker is ˜15 Å.
22. The capture agent of claim 1 , wherein the linker comprises PEG 3 .
23. The capture agent of claim 1 , wherein the linker comprises a peptide.
24. The capture agent of claim 23 , wherein the peptide comprises glycine.
25. A method for detecting IL-17A, IL-17F, or both IL-17A and IL-17F in a biological sample, the method comprising the steps of:
contacting the biological sample with one or more of the capture agents of claim 16 ;
allowing the capture agents to bind to the IL-17A, IL-17F or both IL-17A and IL-17F; and
detecting the bound capture agents via the detectable moiety.
26. The capture agent of claim 1 , wherein the first epitope comprises the amino acid sequence PNSEDKNFPRTVMVNL (SEQ ID NO:43).
27. The capture agent of claim 1 , wherein the first ligand has structure
wherein X represents the rest of the capture agent.
28. The capture agent of claim 6 , wherein the second ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64)vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), and rsynk (SEQ ID NO:76).
29. The capture agent of claim 6 , wherein the second ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).
30. The capture agent of claim 7 , wherein the first ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).
31. The capture agent of claim 1 , wherein the first ligand comprises a first peptide having an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), rkhyh (SEQ ID NO:46), rrATS (SEQ ID NO:47), rrAQS (SEQ ID NO:48), rrats (SEQ ID NO:49), and rraqs (SEQ ID NO:50),
wherein the second ligand comprises a second peptide having an amino acid sequence selected from the group consisting of 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), RSYNK (SEQ ID NO:28), kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64), vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), rsynk (SEQ ID NO:76), rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).
32. The capture agent of claim 31 , wherein the first ligand comprises an amino acid sequence selected from the group consisting of rrATS (SEQ ID NO:47), rrAQS (SEQ ID NO:48), rrats (SEQ ID NO:49), and rraqs (SEQ ID NO:50).
33. The capture agent of claim 32 , wherein the second ligand comprises an amino acid sequence selected from the group consisting of kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64)vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), and rsynk (SEQ ID NO:76).
34. The capture agent of claim 32 , wherein the second ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).
35. The capture agent of claim 31 , wherein the second ligand comprises an amino acid sequence selected from the group consisting of kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64)vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), and rsynk (SEQ ID NO:76).
36. The capture agent of claim 35 , wherein the first ligand comprises an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).
37. A capture agent for a target, the capture agent comprising two or more ligands covalently linked to each other, wherein the ligands each specifically bind to one of two or more distinct epitopes of the target that are in different locations on the target,
wherein the capture agent comprises a first of the ligands having specific affinity for a first of the epitopes, a second of the ligands having specific affinity for a second of the epitopes, and a linker covalently connecting the first ligand to the second ligand,
wherein the target is IL-17A, IL-17F, or both IL-17A and IL-17F,
wherein the first epitope comprises the amino acid sequence FFQKPES (SEQ ID NO:1) or the amino acid sequence PNSEDKNFPRTVMVNL (SEQ ID NO:43), wherein the second epitope comprises the amino acid sequence NENQRVS (SEQ ID NO:3) or the amino acid sequence PNSEDKNFPRTVMVNL (SEQ ID NO:43),
wherein the first ligand comprises a first peptide having an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), rkhyh (SEQ ID NO:46), 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), RRAQS (SEQ ID NO:10), rrATS (SEQ ID NO:47), rrAQS (SEQ ID NO:48), rrats (SEQ ID NO:49), and rraqs (SEQ ID NO:50),
wherein the second ligand comprises a second peptide having an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64), vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), rsynk (SEQ ID NO:76), rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46),
wherein the first ligand is cyclic, and wherein the second ligand is cyclic.
38. The capture agent of claim 37 , wherein the second ligand comprises an amino acid sequence 80-100% identical to an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).
39. The capture agent of claim 37 , wherein the first ligand comprises a first peptide having an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), rkhyh (SEQ ID NO:46), 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), RRAQS (SEQ ID NO:10), rrATS (SEQ ID NO:47), rrAQS (SEQ ID NO:48), rrats (SEQ ID NO:49), and rraqs (SEQ ID NO:50),
wherein the second ligand comprises a second peptide having an amino acid sequence selected from the group consisting of kYGEV (SEQ ID NO:51), VHkSG (SEQ ID NO:52), QkHGP (SEQ ID NO:53), TkHGP (SEQ ID NO:54), YDLQr (SEQ ID NO:55), YDLTr (SEQ ID NO:56), YDkQr (SEQ ID NO:57), YDkTr (SEQ ID NO:58), kkGWP (SEQ ID NO:59), kLGWP (SEQ ID NO:60), LkGWP (SEQ ID NO:61), rSYNL (SEQ ID NO:62), rSYNk (SEQ ID NO:63), kygev (SEQ ID NO:64), vhksg (SEQ ID NO:65), qkhgp (SEQ ID NO:66), tkhgp (SEQ ID NO:67), ydlqr (SEQ ID NO:68), ydltr (SEQ ID NO:69), ydkqr (SEQ ID NO:70), ydktr (SEQ ID NO:71), kkgwp (SEQ ID NO:72), klgwp (SEQ ID NO:73), lkgwp (SEQ ID NO:74), rsynl (SEQ ID NO:75), rsynk (SEQ ID NO:76), rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46),
wherein the first ligand is cyclic, and wherein the second ligand is cyclic.
40. The capture agent of claim 39 , wherein the second ligand comprises an amino acid sequence selected from the group consisting of rhfrl (SEQ ID NO:44), nrfff (SEQ ID NO:45), and rkhyh (SEQ ID NO:46).