Polypeptides and uses thereof
Provided herein are novel polypeptides, methods for their production and uses thereof.
1. An antibody, wherein a polypeptide chain of the antibody is linked on its C-terminus to a coiled-coil peptide, wherein the coiled-coil peptide is capable of forming a heterodimer with another coiled-coil peptide and comprises the amino acid sequence selected from the group consisting of:
SPEDEIQQLEEEIAQLEQKNAALKEKNQALKYGKG (SEQ ID NO: 10);
SPEDKIAQLKEK NAALKEKNQQLKEKLQALKYG (SEQ ID NO: 11);
SPEDEIQQLEEEIAQLEQKNAALKEKNQALKYG (SEQ ID NO: 12);
SPEDKIAQLKQKIQALKQENQQLEEENAALEYG (SEQ ID NO: 13);
MK LKKIKSV LKKIKSV LKKIKSV LKKIKSV VGER (SEQ ID NO: 1);
LKKIKSV LKKIKSV LKKIKSV LKKIKSV (SEQ ID NO: 5);
LKKIKSV LKKIKSV LKKIKKV LKKIKYV (SEQ ID NO: 6);
LEEIVSE LEEIVSE LEEIVSE LEEIVSE (SEQ ID NO: 7); or
LEEIVSE LEEIVSE LEEIVKE LEEIVYE (SEQ ID NO: 8), and
wherein the antibody further comprises a second coiled-coil peptide forming a heterodimer with the coiled-coil peptide linked to the polypeptide chain of the antibody, and wherein the second coiled-coil peptide comprises the amino acid sequence SPEDEIQALEEENAQLEQENAALEEELAQLEYGK-azidoG (SEQ ID NO: 4); MK-LEEIVSE-LEEIVSE-LEEIV(X 1 )E-LEEIVYE-VGER (SEQ ID NO: 51), where X 1 is lysine linked at N-ε position with —X 2 -L, where X 2 is a linker and L is a ligand; or SPEDEIQALEEENAQLEQENAALEEE LAQLEYG(X 1 )KG (SEQ ID NO: 52), where X 1 is lysine linked at N-ε position with —X 2 -L, where X 2 is a linker and L is a ligand.
2. The antibody of claim 1 , wherein said polypeptide chain is a heavy chain of the antibody.
3. The antibody of claim 1 , wherein the coiled-coil peptide linked to the polypeptide chain of the antibody comprises the amino acid sequence:
SPEDEIQQLEEEIAQLEQKNAALKEKNQALKYGKG (SEQ ID NO: 10);
SPEDKIAQLKEK NAALKEKNQQLKEKLQALKYG (SEQ ID NO: 11);
SPEDEIQQLEEEIAQLEQKNAALKEKNQALKYG (SEQ ID NO: 12); or
SPEDKIAQLKQKIQALKQENQQLEEENAALEYG (SEQ ID NO: 13).
4. The antibody of claim 3 , wherein the coiled-coil peptide linked to the polypeptide chain of the antibody comprises the amino acid sequence of SEQ ID NO: 10.
5. The antibody of claim 1 , wherein the coiled-coil peptide linked to the polypeptide chain of the antibody is linked to the C-terminus of the heavy chain via a peptide linker.
6. The antibody of claim 5 , wherein the linker comprises a recognition amino acid sequence for a peptide ligase.
7. The antibody of claim 6 , wherein the linker comprises a Sortase A recognition sequence.
8. The antibody of claim 7 , wherein the linker comprises the amino acid sequence LPETG (SEQ ID NO: 48) or LPETGGGGG (SEQ ID NO: 50).
9. The antibody of claim 1 , wherein the antibody is an anti-GPR87 antibody or an anti-DEspR antibody.
10. The antibody of claim 1 , wherein the antibody is a monoclonal antibody.
11. The antibody of claim 1 , wherein the second coiled-coil peptide comprises the amino acid sequence SPEDEIQALEEENAQLEQENAALEEE LAQLEYGK-azidoG (SEQ ID NO: 4).
12. The antibody of claim 1 , wherein the second coiled-coil peptide comprises the amino acid sequence SPEDEIQALEEENAQLEQENAALEEE LAQLEYG(X 1 )KG (SEQ ID NO: 52), where X 1 is lysine linked at N-ε position with —X 2 -L, where X 2 is a linker and L is a ligand.
13. The antibody of claim 1 , wherein the ligand is selected from the group consisting of small organic and inorganic molecules, amino acids, peptides, polypeptides, peptidomimetics, glycoproteins, lectins, nucleosides, nucleotides, monosaccharides, disaccharides, trisaccharides, oligosaccharides, polysaccharides, lipopolysaccharides, vitamins, steroids, hormones, and cofactors.
14. The antibody of claim 1 , wherein the ligand is a therapeutic agent, an imaging agent or a detectable label.
15. The antibody of claim 14 , wherein the ligand is an anti-cancer agent.
16. The antibody of claim 1 , wherein the ligand is linked to the second coiled-coil peptide via a linker comprising a cleavable linking group.
17. The antibody of claim 1 , wherein the linker linking the ligand to the second coiled-coil peptide comprises the tripeptide glutamic acid-valine-citrulline (EVC).
18. The antibody of claim 17 , wherein the linker comprises triethyleneglycol-glutamic acid-valine-citrulline-p-aminobenzylcarbamate (PEG 3 -EVC-PABC).
19. The antibody of claim 1 , wherein the coiled-coil peptide linked to the polypeptide chain of the antibody comprises the amino acid sequence GGGGG (SEQ ID NO: 55) at its N-terminus.
20. The antibody of claim 1 , wherein the second coiled-coil peptide comprises the amino acid sequence MK-LEEIVSE-LEEIVSE-LEEIV(X 1 )E-LEEIVYE-VGER (SEQ ID NO: 51), where X 1 is lysine linked at N-ε position with —X 2 -L, where X 2 is a linker and L is a ligand.
21. An antibody, wherein a heavy chain of the antibody is linked on its C-terminus to a coiled-coil peptide, wherein the coiled-coil peptide is capable of forming a heterodimer with another coiled-coil peptide and comprises the amino acid sequence SEQ ID NO: 10, wherein the coiled-coil peptide is linked to the C-terminus of the heavy chain via a peptide linker comprising the amino acid sequence of SEQ ID NO: 55, wherein the antibody is conjugated with a ligand, and wherein the ligand is covalently linked with a coiled-coil peptide comprising the amino acid sequence of SEQ ID NO: 52.
22. A pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.
23. A method of delivering at least one ligand to a cell, the method comprising providing to the cell the antibody of claim 1 .