Modified J-chain
The present invention concerns modified recombinant J-chain polypeptides, binding molecules, such as antibodies comprising the same, and their uses.
1. A polymeric antibody that specifically binds to an antigen, the polymeric antibody comprising at least two immunoglobulin monomers and a modified J-chain, wherein the modified J-chain comprises an extraneous binding moiety introduced into a native J-chain comprising SEQ ID NO: 1, wherein the extraneous binding moiety is introduced into the native J-chain at or within 10 residues from the C- or N-terminus of the native J-chain, and wherein each immunoglobulin monomer comprises (i) a Cμ4 and an IgM tail-piece (μtp) domain, or (ii) a Cα3 and an IgA tail-piece (αtp) domain.
2. The polymeric antibody of claim 1 , wherein the extraneous binding moiety is introduced into the native J-chain by indirect fusion through a peptide linker.
3. The polymeric antibody of claim 2 , wherein the peptide linker is located at or around the C- and/or N-terminus of the binding moiety.
4. The polymeric antibody of claim 2 , wherein the extraneous binding moiety is introduced into the native J-chain at the C-terminus of the native J-chain.
5. The polymeric antibody of claim 2 , wherein the extraneous binding moiety is introduced into the native J-chain at the N-terminus of the native J-chain.
6. The polymeric antibody of claim 2 , wherein the peptide linker is about 10 to 20 amino acids long.
7. The polymeric antibody of claim 2 , wherein the peptide linker is 15 amino acids long.
8. The polymeric antibody of claim 2 , further comprising an additional extraneous binding moiety, wherein the additional extraneous binding moiety is a polypeptide introduced into the native J-chain by direct or indirect fusion.
9. The polymeric antibody of claim 1 , wherein the extraneous binding moiety is selected from the group consisting of an antibody, an antigen-binding fragment of an antibody, an antibody-drug conjugate, an antibody-like molecule, an antigen-binding fragment of an antibody-like molecule, a soluble protein, a ligand, a receptor, a virus-like particle, a protein toxin, an enzyme, a darpin, a fibronectin domain, an adnectin, and a knottin.
10. The polymeric antibody of claim 9 , wherein the extraneous binding moiety is an antigen-binding fragment of an antibody selected from the group consisting of an F(ab′)2, an F(ab)2, an Fab′, an Fab, an Fv, an scFv, and a single domain antibody.
11. The polymeric antibody of claim 10 , wherein the antigen-binding fragment is an scFv.
12. The polymeric antibody of claim 1 , wherein the extraneous binding moiety binds to an effector cell selected from the group consisting of a T-cell, a natural killer (NK) cell, a macrophage, and a neutrophil.
13. The polymeric antibody of claim 12 , wherein the effector cell is a T-cell.
14. The polymeric antibody of claim 13 , wherein the extraneous binding moiety binds to CD3ε on the T-cell.
15. The polymeric antibody of claim 13 , wherein the extraneous binding moiety binds to CD3ε on a T-cell.
16. The polymeric antibody of claim 1 , which comprises five immunoglobulin monomers to form a pentamer, wherein each immunoglobulin monomer comprises a μtp domain and a Cμ4 domain.
17. The polymeric antibody of claim 16 , wherein each immunoglobulin monomer each further comprise a Cμ1 domain, a Cμ2 domain, and a Cμ3 domain.
18. The polymeric antibody of claim 16 , wherein each immunoglobulin monomer comprises one or more heavy chain constant region domains of a non-IgM isotype.
19. The polymeric antibody of claim 18 , wherein the non-IgM isotype is IgG.
20. The polymeric antibody of claim 1 , which comprises two immunoglobulin monomers to form a dimer, wherein each immunoglobulin monomer comprises an αtp domain and a Cα3 domain.
21. The polymeric antibody of claim 20 , wherein each immunoglobulin monomer further comprises a Cα1 domain and a Cα2 domain.
22. The polymeric antibody of claim 20 , wherein each immunoglobulin monomer comprises one or more heavy chain constant region domains of a non-IgA isotype.
23. The polymeric antibody of claim 22 , wherein the non-IgA isotype is IgG.
24. The polymeric antibody of claim 1 , wherein the extraneous binding moiety is an scFv.
25. The polymeric antibody of claim 24 , which comprises five immunoglobulin monomers to form a pentamer, wherein each immunoglobulin monomer comprises a μtp domain and a Cμ4 domain.
26. The polymeric antibody of claim 25 , wherein each immunoglobulin monomer each further comprise a Cμ1 domain, a Cμ2 domain, and a Cμ3 domain.
27. The polymeric antibody of claim 25 , wherein each the immunoglobulin monomer comprises one or more heavy chain constant region domains of a non-IgM isotype.
28. The polymeric antibody of claim 27 , wherein the non-IgM isotype is IgG.