IgM Fc and J-chain mutations that affect IgM serum half-life
This disclosure provides an IgM antibody or IgM-like antibody comprising a variant J-chain and/or variant IgM heavy chain constant regions that can confer increased serum half-life upon the antibody.
1. An IgM antibody with enhanced serum half-life comprising five bivalent antibody binding units and a variant J-chain,
wherein each binding unit comprises two IgM heavy chain constant regions, each associated with a single chain Fv (scFv), single domain variable region (VHH), or heavy chain variable region (VH),
wherein the variant J-chain comprises an alanine (A), or arginine (R) substitution at a position corresponding to amino acid Y102 of SEQ ID NO: 2, an alanine (A) substitution at a position corresponding to amino acid N49 of SEQ ID NO: 2, or an alanine (A), or arginine (R) substitution at a position corresponding to amino acid Y102 of SEQ ID NO: 2 and an alanine (A) substitution at a position corresponding to amino acid N49 of SEQ ID NO: 2, wherein the variant J-chain can affect serum half-life of the IgM antibody; and
wherein the IgM antibody exhibits an increased serum half-life upon administration to a subject animal relative to a reference IgM antibody that is identical except for the amino acid substitution in the variant J-chain, and is administered in the same way to the same animal species.
2. The IgM antibody of claim 1 , wherein the variant J-chain comprises an alanine (A), or arginine (R) substitution at a position corresponding to Y102 of SEQ ID NO: 2.
3. The IgM antibody of claim 2 , wherein the amino acid corresponding to Y102 of SEQ ID NO: 2 is substituted with alanine (A).
4. The IgM antibody of claim 3 , wherein the J-chain is a variant human J-chain and comprises the amino acid sequence SEQ ID NO: 3.
5. The IgM antibody of claim 1 , wherein the variant J-chain comprises an alanine (A) substitution at the position corresponding to N49 of SEQ ID NO: 2.
6. The IgM antibody of claim 1 , wherein the IgM heavy chain constant regions are variant IgM heavy chain constant regions comprising an alanine (A) substitution at a position corresponding to amino acid 5401 of SEQ ID NO: 12, an alanine (A) substitution at a position corresponding to amino acid E402 of SEQ ID NO: 12, or alanine (A) substitutions at positions corresponding to amino acids 5401 and E402 of SEQ ID NO: 12, wherein the variant IgM heavy chain constant regions can affect serum half-life of the IgM antibody, and wherein the IgM antibody exhibits a further increased serum half-life upon administration to a subject animal relative to a reference IgM antibody that is identical except for the amino acid substitutions in the IgM heavy chain constant regions, and is administered in the same way to the same animal species.
7. An IgM antibody with enhanced serum half-life comprising five or six bivalent antibody binding units,
wherein each binding unit comprises two variant IgM heavy chain constant regions, each associated with a single chain Fv (scFv), single domain variable region (VHH), or heavy chain variable region (VH),
wherein the variant IgM heavy chain constant regions each comprise an alanine (A) substitution at a position corresponding to amino acid 5401 of SEQ ID NO: 12, an alanine (A) substitution at a position corresponding to amino acid E402 of SEQ ID NO: 12, or alanine (A) substitutions at positions corresponding to amino acids 5401 and E402 of SEQ ID NO: 12, wherein the variant IgM heavy chain constant regions can affect serum half-life of the IgM antibody; and
wherein the IgM antibody exhibits an increased serum half-life upon administration to a subject animal relative to a reference IgM antibody that is identical except for the amino acid substitution, and is administered in the same way to the same animal species.
8. The IgM antibody of claim 1 , wherein the increased serum half-life comprises an increased alpha half-life (t 1/2 α), an increased beta half-life (t 1/2 β), or an increased t 1/2 α and an increased t 1/2 β.
9. The IgM antibody of claim 8 , which further exhibits an increased peak plasma concentration (Cmax), an increased area under the curve (AUC), a modified clearance time, or a combination thereof relative to the reference antibody.
10. The IgM antibody of claim 1 , wherein the IgM heavy chain constant regions each comprise a Cμ4 domain and an IgM tailpiece (tp) domain.
11. The IgM antibody of claim 10 , wherein the IgM heavy chain constant regions each further comprise a Cμ3 domain, a Cμ2 domain, a Cμ1 domain, or a combination thereof.
12. The IgM antibody of claim 1 , wherein the IgM heavy chain constant regions are each associated with a heavy chain variable region (VH).
13. The IgM antibody of claim 12 , wherein each binding unit further comprises two light chain constant regions each associated with a light chain variable region (VL).
14. The IgM antibody of claim 13 , wherein the light chain constant regions are kappa or lambda light chain constant regions.
15. The IgM antibody of claim 1 , wherein the variant J-chain further comprises one or more heterologous polypeptides directly or indirectly fused to the variant J-chain.
16. The IgM antibody of claim 15 , wherein the one or more heterologous polypeptides is/are fused to the variant J-chain via a peptide linker.
17. The IgM antibody of claim 16 , wherein the peptide linker comprises at least 5 amino acids, but no more than 25 amino acids.
18. The IgM antibody of claim 15 , wherein the one or more heterologous polypeptides is/are fused to the N-terminus of the variant J-chain, the C-terminus of the variant J-chain, or wherein heterologous polypeptides are fused to both the N-terminus and C-terminus of the variant J-chain, wherein the heterologous polypeptides can be the same or different.
19. The IgM antibody of claim 15 , wherein at least one heterologous polypeptide comprises a binding domain comprising an antibody or antigen-binding fragment thereof.
20. The IgM antibody of claim 19 , wherein the antigen-binding fragment comprises a Fab, a Fab′, a F(ab′)2, a Fd, a Fv, a single-chain Fv (scFv), a disulfide-linked Fv (sdFv), or a combination thereof.
21. The IgM antibody of claim 20 , wherein the antigen-binding fragment is a scFv.
22. The IgM antibody of claim 21 , wherein at least one heterologous polypeptide can specifically bind to CD3ε.
23. A composition comprising the IgM antibody of claim 1 , and a pharmaceutically acceptable carrier.
24. The IgM antibody of claim 7 , wherein the variant IgM heavy chain constant regions comprise an alanine (A) substitution at the position corresponding to amino acid S401 of SEQ ID NO: 12.
25. The IgM antibody of claim 7 , wherein the variant IgM heavy chain constant regions comprise an alanine (A) substitution at the position corresponding to E402 of SEQ ID NO: 12.
26. The IgM antibody of claim 7 , wherein the variant IgM heavy chain constant regions are variant human IgM heavy chain constant regions and comprise the amino acid sequence SEQ ID NO: 13 or SEQ ID NO: 14.
27. The IgM antibody of claim 7 , wherein the increased serum half-life comprises an increased alpha half-life (t 1/2 α), an increased beta half-life (t 1/2 β), or an increased t 1/2 α and an increased t 1/2 β.
28. A composition comprising the IgM antibody of claim 7 , and a pharmaceutically acceptable carrier.