Heterodimeric Fc regions, binding molecules comprising same, and methods relating thereto
The present invention features inter alia polypeptides comprising heterodimeric Fc regions. In addition, the instant invention provides methods for treating or preventing a disease or disorder in subject by administering the polypeptides of the invention to said subject.
1. A binding molecule comprising:
(i) a binding moiety, wherein the binding moiety comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL),
(ii) a first Fc polypeptide chain comprising a first CH2 domain operably linked to the N-terminus of a CL moiety (CH2-CL), and
(iii) a second Fc polypeptide chain comprising a second CH2 domain operably linked to the N-terminus of a CH1 moiety (CH2-CH1),
wherein the binding moiety is linked to one or both of the first and second Fc polypeptide chains,
wherein said first and second Fc polypeptide chains form a heterodimeric Fc region via CL moiety:CH1 moiety heterodimerization,
wherein the CL moiety comprises an amino acid sequence selected from the group consisting of amino acids:
126-233 of SEQ ID NO:28,
126-233 of SEQ ID NO:29,
126-233 of SEQ ID NO:30,
126-235 of SEQ ID NO:31,
126-235 of SEQ ID NO:32,
126-235 of SEQ ID NO:33,
126-237 of SEQ ID NO:34,
126-237 of SEQ ID NO:35,
126-237 of SEQ ID NO:36, and
153-260 of SEQ ID NO:46, and
wherein the CH1 moiety comprises an amino acid sequence selected from the group consisting of amino acids:
126-228 of SEQ ID NO:37,
126-224 of SEQ ID NO:38,
126-224 of SEQ ID NO:39,
126-224 of SEQ ID NO:40,
126-225 of SEQ ID NO:41,
126-226 of SEQ ID NO:42,
126-226 of SEQ ID NO:43,
126-226 of SEQ ID NO:44,
126-227 of SEQ ID NO:45,
406-504 of SEQ ID NO:47,
406-504 of SEQ ID NO:48, and
406-504 of SEQ ID NO:49.
2. The binding molecule of claim 1 , wherein said first Fc polypeptide chain further comprises an Fc moiety operably linked to the C-terminus of the CL moiety (CH2-CL-Fc) and said second Fc polypeptide chain further comprises an Fc moiety operably linked to the C-terminus of the CH1 moiety (CH2-CH1-Fc).
3. The binding molecule of claim 1 , wherein the CH1 moiety comprises an amino acid sequence selected from the group consisting of amino acids 406-504 of SEQ ID NO:47, amino acids 406-504 of SEQ ID NO:48, and amino acids 406-504 of SEQ ID NO:49, and wherein the CL moiety comprises the amino acid sequence of amino acids 153-260 of SEQ ID NO:46.
4. The binding molecule of claim 1 , wherein both the first and second Fc polypeptide chains are aglycosylated.
5. The binding molecule of claim 1 , wherein said binding moiety is selected from the group consisting of an scFv, an Fab, and a diabody.
6. The binding molecule of claim 1 , wherein the binding moiety specifically binds an antigen selected from the group consisting of CD20, CD2, CD3, CD5, CD6, CD7, MAGE-1, MAGE-3, MUC-1, HPV16, HPV E6, HPV E7, TAG-72, CEA, α-Lewis y , L6-Antigen, CD19, CD22, CD23, CD25, CD30, CD33, CD37, CD44, CD52, CD56, mesothelin, PSMA, HLA-DR, EGF receptor, VEGF, VEGF receptor, Cripto antigen, HER2 receptor, c-MET, RON, TNFR2, HVEM, CD27, CD40, 4-1BB, OX40, and GITR.
7. A multivalent binding molecule comprising:
(i) a first binding moiety comprising a first immunoglobulin heavy chain variable domain (VH) and a first immunoglobulin light chain variable domain (VL), the first binding moiety operably linked or fused to a first Fc polypeptide chain comprising a first CH2 domain operably linked to the N-terminus of a CL moiety (CH2-CL), and
(ii) a second binding moiety comprising a second immunoglobulin heavy chain variable domain (VH) and a second immunoglobulin light chain variable domain (VL), the second binding moiety operably linked or fused to a second Fc polypeptide chain comprising a second CH2 domain operably linked to the N-terminus of a CH1 moiety (CH2-CH1),
wherein said first and second Fc polypeptide chains form a heterodimeric Fc region via CL moiety:CH1 moiety heterodimerization.
8. The binding molecule of claim 7 , wherein heterodimerization is stabilized by one or more interchain disulfide bonds between said CH2-CL and said CH2-CH1.
9. The binding molecule of claim 7 , wherein said first Fc polypeptide chain further comprises a hinge region.
10. The binding molecule of claim 7 , wherein said second Fc polypeptide chain further comprises a hinge region.
11. The binding molecule of claim 7 , wherein said first and second CH2 domains are of the IgG1sotype.
12. A composition comprising a population of the binding molecule of claim 7 , wherein at least 90% of binding molecules in the population are in heterodimeric form.
13. A pharmaceutical composition comprising the binding molecule of claim 7 , and a pharmaceutically acceptable carrier.
14. The binding molecule of claim 1 , wherein the CH2-CL comprises amino acids 43-260 of SEQ ID NO:46 and the CH2-CH1 comprises amino acids 296-504 of SEQ ID NO:47.
15. The binding molecule of claim 1 , wherein the CH2-CL comprises amino acids 43-260 of SEQ ID NO:46 and the CH2-CH1 comprises amino acids 296-504 of SEQ ID NO:48.
16. The binding molecule of claim 1 , wherein the CH2-CL comprises amino acids 43-260 of SEQ ID NO:46 and the CH2-CH1 comprises amino acids 296-504 of SEQ ID NO:49.
17. The binding molecule of claim 1 , wherein the first and second CH2 domains have the same amino acid sequence.
18. The binding molecule of claim 1 , wherein the first and second CH2 domains are either an IgG1 CH2 domain or an IgG4 CH2 domain.
19. The binding molecule of claim 1 , wherein the CH2-CL comprises an amino acid sequence selected from the group consisting of:
16-233 of SEQ ID NO:28,
16-233 of SEQ ID NO:29,
16-233 of SEQ ID NO:30,
16-235 of SEQ ID NO:31,
16-235 of SEQ ID NO:32,
16-235 of SEQ ID NO:33,
16-237 of SEQ ID NO:34,
16-237 of SEQ ID NO:35,
16-237 of SEQ ID NO:36, and
43-260 of SEQ ID NO:46.
20. The binding molecule of claim 1 , wherein the CH2-CH1 comprises an amino acid sequence selected from the group consisting of:
16-228 of SEQ ID NO:37,
16-224 of SEQ ID NO:38,
16-224 of SEQ ID NO:39,
16-224 of SEQ ID NO:40,
16-225 of SEQ ID NO:41,
16-226 of SEQ ID NO:42,
16-226 of SEQ ID NO:43,
16-226 of SEQ ID NO:44,
16-227 of SEQ ID NO:45,
296-509 of SEQ ID NO:47,
296-509 of SEQ ID NO:48, and
296-509 of SEQ ID NO:49.
21. The binding molecule of claim 1 , wherein the first and second Fc polypeptide chains each further comprise a hinge sequence set forth in SEQ ID NO:59 or SEQ ID NO:64.
22. The binding molecule of claim 17 , wherein the first and second Fc polypeptide chains each further comprise a hinge sequence set forth in SEQ ID NO:59 or SEQ ID NO:64.
23. The binding molecule of claim 1 , comprising two binding moieties.
24. A binding molecule comprising:
(i) a binding moiety, wherein the binding moiety is an scFv,
(ii) a first Fc polypeptide chain comprising a first CH2 domain operably linked to the N-terminus of a CL moiety (CH2-CL), and
(iii) a second Fc polypeptide chain comprising a second CH2 domain operably linked to the N-terminus of a CH1 moiety ((CH2-CH1),
wherein the binding moiety is linked to one of the first and second Fc polypeptide chains,
wherein the first and second Fc polypeptide chains form a heterodimeric Fc region via CL moiety:CH1 moiety heterodimerization,
wherein the CL moiety comprises an amino acid sequence selected from the group consisting of amino acids:
126-233 of SEQ ID NO:28,
126-233 of SEQ ID NO:29,
126-233 of SEQ ID NO:30,
126-235 of SEQ ID NO:31,
126-235 of SEQ ID NO:32,
126-235 of SEQ ID NO:33,
126-237 of SEQ ID NO:34,
126-237 of SEQ ID NO:35,
126-237 of SEQ ID NO:36, and
153-260 of SEQ ID NO:46, and
wherein the CH1 moiety comprises an amino acid sequence selected from the group consisting of amino acids:
126-228 of SEQ ID NO:37,
126-224 of SEQ ID NO:38,
126-224 of SEQ ID NO:39,
126-224 of SEQ ID NO:40,
126-225 of SEQ ID NO:41,
126-226 of SEQ ID NO:42,
126-226 of SEQ ID NO:43,
126-226 of SEQ ID NO:44,
126-227 of SEQ ID NO:45,
406-504 of SEQ ID NO:47,
406-504 of SEQ ID NO:48, and
406-504 of SEQ ID NO:49.
25. The binding molecule of claim 24 , wherein the CL moiety comprises the amino acid sequence of amino acids 153-260 of SEQ ID NO:46 and the CH1 moiety comprises the amino acid sequence of amino acids 406-504 of SEQ ID NO:47.
26. The binding molecule of claim 24 , wherein the CL moiety comprises the amino acid sequence of amino acids 153-260 of SEQ ID NO:46 and the CH1 moiety comprises the amino acid sequence of amino acids 406-504 of SEQ ID NO:48.
27. The binding molecule of claim 24 , wherein the CL moiety comprises the amino acid sequence of amino acids 153-260 of SEQ ID NO:46 and the CH1 moiety comprises the amino acid sequence of amino acids 406-504 of SEQ ID NO:49.
28. The binding molecule of claim 24 , wherein the first and second CH2 domains are each either an IgG1 CH2 domain or an IgG4 CH2 domain.
29. A binding molecule comprising:
(i) a binding moiety, wherein the binding moiety comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL),
(ii) a first Fc polypeptide chain comprising a first CH3 domain operably linked to the N-terminus of a CL moiety (CH3-CL), and
(iii) a second Fc polypeptide chain comprising a second CH3 domain operably linked to the N-terminus of a CH1 moiety (CH3-CH1),
wherein the binding moiety is linked to one or both of the first and second Fc polypeptide chains,
wherein said first and second Fc polypeptide chains form a heterodimeric Fc region via CL moiety:CH1 moiety heterodimerization,
wherein the CL moiety comprises an amino acid sequence selected from the group consisting of amino acids:
126-233 of SEQ ID NO:28,
126-233 of SEQ ID NO:29,
126-233 of SEQ ID NO:30,
126-235 of SEQ ID NO:31,
126-235 of SEQ ID NO:32,
126-235 of SEQ ID NO:33,
126-237 of SEQ ID NO:34,
126-237 of SEQ ID NO:35,
126-237 of SEQ ID NO:36, and
153-260 of SEQ ID NO:46, and
wherein the CH1 moiety comprises an amino acid sequence selected from the group consisting of amino acids:
126-228 of SEQ ID NO:37,
126-224 of SEQ ID NO:38,
126-224 of SEQ ID NO:39,
126-224 of SEQ ID NO:40,
126-225 of SEQ ID NO:41,
126-226 of SEQ ID NO:42,
126-226 of SEQ ID NO:43,
126-226 of SEQ ID NO:44,
126-227 of SEQ ID NO:45,
406-504 of SEQ ID NO:47,
406-504 of SEQ ID NO:48, and
406-504 of SEQ ID NO:49.
30. The binding molecule of claim 29 , wherein the first and second Fc polypeptide chains further comprise a first CH2 domain [CH2-CH3-CL] and second CH2 domain [CH2-CH3-CH1], respectively.
31. The binding molecule of claim 29 , wherein the first and second CH3 domains are each an IgG4 CH3 domain.
32. The binding molecule of claim 30 , wherein the CH2-CH3-CL comprises an amino acid sequence selected from the group consisting of:
40-366 of SEQ ID NO:69,
40-371 of SEQ ID NO:71, and
43-369 of SEQ ID NO:73.
33. The binding molecule of claim 30 , wherein the CH2-CH3-CH1 comprises an amino acid sequence selected from the group consisting of:
293-613 of SEQ ID NO:70,
293-618 of SEQ ID NO:72, and
296-616 of SEQ ID NO:74.