Bispecific antibodies with alternatively matched interchain cysteines and uses thereof
AssigneePhanes Therapeutics, Inc.
InventorsJack Chongyang Li, Minghan Wang, Hui Zou, Haiqun Jia
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Abstract
Engineered bispecific antibodies with shifted interchain disulfide bond on one arm while maintaining the native interchain disulfide bond on the second arm are described. Also described are anti-CD47/FRα bispecific antibodies and antigen-binding fragments thereof. Also described are nucleic acids encoding the antibodies, compositions comprising the antibodies, and methods of producing the antibodies and using the antibodies for treating or preventing diseases, such as cancer and/or associated complications.
Claims19
An isolated bispecific antibody or antigen-binding fragment thereof comprising:
- a. a first heavy chain, H1;
- b. a second heavy chain, H2;
- c. a first light chain, L1;
- and
- d. a second light chain, L2;
- wherein H1 and L1 form a first arm comprising a first antigen-binding domain that specifically binds a first antigen, wherein the first antigen-binding domain comprises a VH region and a VL region, and
- wherein H2 and L2 form a second arm comprising a second antigen-binding domain that specifically binds a second antigen, wherein the second antigen-binding domain comprises a VH region and a VL region, wherein
- (a) H1 comprises a CH1 region of human IgG1, IgG2, IgG3, or IgG4;
- and
- (b) L1 comprises a CL region of a human kappa light chain or a human lambda light chain;
- wherein the CH1 region sequence of H1 is selected from the group consisting of SEQ ID NO:15, 21, 22, and 23;
- the CL region sequence of L1 is selected from SEQ ID NO:19 or 24;
- and wherein the CH1 and CL regions further comprise amino acid substitutions selected from:
- (1) K133C and C220X in CH1, and F209C and C214X in CL;
- (2) K133C and C220X in CH1, and K207C and C214X in CL;
- (3) R133C and C131X in CH1, and K207C and C214X in CL;
- (4) R133C and C131X in CH1, and F209C and C214X in CL;
- (5) R133C and C131X in CH1, and V209C and C214X in CL;
- or
- (6) K133C and C220X in CH1, and V209C and C214X in CL;
- wherein X is selected from S, A or G;
- wherein the second arm comprising H2 and L2 does not comprise the amino acid substitutions of the first arm comprising H1 and L1, maintaining its native interchain disulfide bond, and wherein EU numbering is used for the CH1 and CL regions.
An isolated bispecific antibody or antigen-binding fragment thereof comprising:
- a. a first heavy chain, H1;
- b. a second heavy chain, H2;
- c. a first light chain, L1;
- and
- d. a second light chain, L2;
- wherein H1 and L1 form a first arm comprising a first antigen-binding domain that specifically binds a first antigen, wherein the first antigen-binding domain comprises a VH region and a VL region, and
- wherein H2 and L2 form a second arm comprising a second antigen-binding domain that specifically binds a second antigen, wherein the second antigen-binding domain comprises a VH region and a VL region, wherein
- (a) H1 comprises a CH1 region of human IgG1, IgG2, IgG3, or IgG4 and a heavy chain variable region (VH region);
- and
- (b) L1 comprises a CL region of a human kappa light chain or a human lambda light chain and a light chain variable region (VL region);
- wherein the CH1 region sequence of H1 is selected from the group consisting of SEQ ID NO:
- 15, 21, 22, and 23, the VH region sequence of H1 is SEQ ID NO:13, the CL region sequence of L1 is selected from SEQ ID NO:19 or 24, and the VL region sequence of L1 is SEQ ID NO:17;
- and
- wherein the CH1, VH1, CL, and VL regions further comprise amino acid substitutions selected from:
- (1) C220X in CH1, G44C in VH, C214X in CL, and G101C in VL;
- or
- (2) C131X in CH1, G44C in VH, C214X in CL, and G101C in VL;
- wherein X is selected from S, A or G;
- wherein the second arm comprising H2 and L2 does not comprise the amino acid substitutions of the first arm comprising H1 and L1, maintaining its native interchain disulfide bond, and wherein EU numbering is used for the CH1, VH, CL, and VL regions.
Dependencies are inferred from the claim text.