Antibodies or fusion proteins multimerized via cysteine mutation and a mu tailpiece
The invention provides constant regions incorporating a cysteine mutation and linked to a μ tailpiece and antibodies or fusion proteins incorporating the same. The constant regions include at least CH2 and CH3 regions of an IgG heavy chain constant region including a cysteine mutation and μ tailpiece. Antibodies or fusion proteins incorporating the constant regions gains the ability to form multivalent complexes, e.g., pentameric or hexameric structures. Antibodies or fusion proteins incorporating the constant regions also retain IgG properties including specific binding to protein G, which facilitates purification and may exhibit pH-dependent FcRn binding, which is associated with a relatively long in vivo half-life. Depending on the isotype and subtype, the nature of the antigen and presence of an additional IgG hinge domain, such antibodies or fusion proteins may also have properties of specific binding to protein A, and effector functions such as ADCC, CDC and opsonization.
1. An antibody comprising IgG hinge, CH2 and CH3 regions in N- to C-terminal order, wherein position 279, 285 or 287 by EU numbering in the CH2 region is mutated to a cysteine residue and the CH3 region is linked to a μ tailpiece at its C-terminus, wherein units of the antibody can multimerize via disulfide bonding between cysteines at the mutated position in different units and between μ tailpieces in different units.
2. The antibody of claim 1 further comprising an IgG CH1 region N-terminal to the hinge.
3. The antibody of claim 1 that specifically binds to a Death Receptor family protein.
4. The antibody of claim 3 , wherein the Death Receptor family protein is DR4 or DR5.
5. The antibody of claim 1 , further comprising a human IgG CH1 region N-terminal to the hinge.
6. The antibody of claim 1 that specifically binds to a TNF receptor family protein.
7. An antibody comprising human IgG1, IgG2 or IgG4 hinge, CH2 and CH3 regions in N- to C-terminal order, wherein position 279, 285 or 287 by EU numbering in the CH2 region is mutated to a cysteine residue and the CH3 region is linked to a μ tailpiece at its C-terminus, wherein units of the antibody can multimerize via disulfide bonding between cysteines at the mutated position in different units and between μ tailpieces in different units.
8. The antibody of claim 1 , which is a single-chain antibody comprising a single-chain Fv linked to the heavy chain constant region.
9. The antibody of claim 8 , which is a component of a multi-specific antibody comprising a plurality of the single-chain antibodies, wherein the single chain Fvs of the plurality have different VH regions, and the plurality of single-chain antibodies are complexed in the multispecific antibody via disulfide bonding between cysteines at the mutated position in different units and between μ tailpieces in different units.
10. The antibody of claim 9 , wherein the single chain Fvs have the same VL region.
11. The antibody of claim 1 , which specifically binds protein G, specifically binds protein A, exhibits ADCC, CDC and/or opsonization.
12. The antibody of claim 11 , wherein a CH1 region, if present, and the hinge region, and CH2 and CH3 regions are human IgG1 regions, and the antibody specifically binds protein G, and specifically binds protein A.
13. The antibody of claim 12 that exhibits ADCC, CDC and opsonizaton.
14. The antibody of claim 1 , wherein a CH1 region if present, and the hinge, CH2 and CH3 regions are human IgG2 or IgG4 regions and the antibody specifically binds protein G and specifically binds protein A.
15. The antibody of claim 1 that is a component of a multispecific complex comprising antibodies complexed by disulfide bonding between the cysteines at the mutated position and between μ tailpieces in different units.
16. The antibody of claim 1 , wherein the antibody is a humanized, chimeric, veneered or human antibody.
17. The antibody of claim 1 that specifically binds the extracellular domain of a receptor.
18. The antibody of claim 17 , which is an antibody that specifically binds to CD79a, CD30, DR5 or DR4.
19. The antibody of claim 17 , which is an antibody that specifically binds to CD40, OX40, 4-1BB, GITR or CD27.
20. A pharmaceutical composition comprising an antibody as defined in claim 1 and a pharmaceutically acceptable carrier.
21. An antibody comprising human IgG hinge, CH2 and CH3 regions in N- to C-terminal order, wherein position 309 in the CH2 region is mutated to a cysteine residue and position 310 is histidine and the CH3 region is linked to a μ tailpiece at its C-terminus, wherein units of the antibody can multimerize via disulfide bonding between cysteines at the mutated position in different units and between μ tailpieces in different units.