Bivalent antibodies masked by coiled coils
The invention provides bivalent antibodies including two light and heavy chain pairs. The N-termini of one or both light and heavy chain pairs are linked via linkers comprising a protease cleavage site to coiled-coil forming peptides that associate to form a coiled coil reducing binding affinity of at least one light-heavy chain pair to a target.
1. A bivalent antibody comprising two light and heavy chain pairs, wherein the N-termini of the light and heavy chains of at least one of the pairs are linked, via linkers comprising a protease cleavage site, to coiled-coil forming peptides that associate to form a coiled coil reducing binding affinity of the antibody to a target relative to the binding affinity of the antibody in naked form; wherein (a) a peptide comprising the sequence QGASTSVDELQAEVDQLEDENYALKTKVAQLRKKVEKLGSIPVSLRSG (SEQ ID NO: 34) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the light chain and a peptide comprising the sequence QGASTTVAQLEEKVKTLRAENYELKSEVQRLEEQVAQLGSIPVSLRSG (SEQ ID NO: 31) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the heavy chain; or (b) a peptide comprising the sequence QGASTTVAQLEEKVKTLRAENYELKSEVQRLEEQVAQLGSIPVSLRSG (SEQ ID NO: 31) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the light chain and a peptide comprising the sequence QGASTSVDELQAEVDQLEDENYALKTKVAQLRKKVEKLGSIPVSLRSG (SEQ ID NO: 34) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the heavy chain.
2. The bivalent antibody of claim 1 , wherein the light and heavy chains of both of the pairs are linked via the linkers comprising the protease cleavage site to the coiled-coil forming peptides that associate to form a coiled coil reducing binding affinity of the antibody to the target.
3. The bivalent antibody of claim 1 , conjugated to a cytotoxic or cytostatic drug.
4. The bivalent antibody of claim 3 , conjugated to a cytotoxic or cytostatic drug via a cysteine residue of the bivalent antibody.
5. The bivalent antibody of claim 1 , wherein the two light and heavy chain pairs are the same.
6. The bivalent antibody of claim 1 , wherein the two light and heavy chain pairs are different.
7. The bivalent antibody of claim 6 , wherein the two light and heavy chain pairs have specificity for different targets.
8. The bivalent antibody of claim 6 , wherein the two light and heavy chain pairs have specificity for the same target.
9. The bivalent antibody of claim 7 , wherein one and only one of the pairs are linked via the linkers comprising the protease cleavage site to the coiled-coil forming peptides that associate to form a coiled coil reducing binding affinity of the antibody to the target.
10. The bivalent antibody of claim 1 , wherein the heavy chain comprises a constant region comprising CH1, hinge, CH2 and CH3 regions.
11. The bivalent antibody of claim 1 , wherein the target or one of the targets is any of CD19, CD30, LIV-1, CD70, or CD74.
12. The bivalent antibody of claim 1 , wherein the binding is reduced 100-1000 fold relative to that of the antibody in naked form.
13. The bivalent antibody of claim 1 , wherein the binding is reduced 100-500 fold relative to that of the antibody in naked form.
14. The bivalent antibody of claim 3 , wherein cytotoxicity of the conjugate is reduced at least 100-1000 fold relative to that of the antibody in naked form.
15. The bivalent antibody of claim 3 , wherein cytotoxicity of the conjugate is reduced 100-500 fold relative to that of the antibody in naked form.
16. The bivalent antibody of claim 1 , wherein multiple copies of the coiled coil forming peptide are linked in tandem to the N-termini of the heavy and light chains.
17. The bivalent antibody of claim 1 , wherein a peptide consisting of the sequence QGASTSVDELQAEVDQLEDENYALKTKVAQLRKKVEKLGSIPVSLRSG (SEQ ID NO: 34) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the light chain and a peptide of consisting of the sequence QGASTTVAQLEEKVKTLRAENYELKSEVQRLEEQVAQLGSIPVSLRSG (SEQ ID NO: 31) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the heavy chain.
18. The bivalent antibody of claim 1 , wherein a peptide consisting of the sequence QGASTTVAQLEEKVKTLRAENYELKSEVQRLEEQVAQLGSIPVSLRSG (SEQ ID NO: 31)) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the light chain and a peptide consisting of the sequence QGASTSVDELQAEVDQLEDENYALKTKVAQLRKKVEKLGSIPVSLRSG (SEQ ID NO: 34) provides the linker comprising the protease cleavage site and the coiled-coil forming peptide linked to the heavy chain.
19. The bivalent antibody of claim 7 , wherein one of the targets is a surface antigen on an immune cell and the other is a surface antigen on a cancer cell.
20. The bivalent antibody of claim 19 , wherein the surface antigen on the immune cell is CD3.