Heterodimeric antibodies that bind fibroblast activation protein
The present invention relates to novel antigen binding domains and heterodimeric antibodies that bind Fibroblast Activation Protein (FAP).
1. A composition comprising a fibroblast activation protein (FAP) binding domain, the FAP binding domain comprising:
a) a variable heavy domain comprising a vhCDR1, vhCDR2, and vhCDR3; and
b) a variable light domain comprising a vlCDR1, vlCDR2, and vlCDR3;
wherein vhCDR1-3 and vlCDR1-3 are selected from the following:
i) a vhCDR1 having SEQ ID NO: 246, a vhCDR2 having SEQ ID NO: 247, a vhCDR3 having SEQ ID NO: 248, a vlCDR1 having SEQ ID NO: 250, a vlCDR2 having SEQ ID NO: 251, and a vlCDR3 having SEQ ID NO: 252;
ii) a vhCDR1 having SEQ ID NO: 230, a vhCDR2 having SEQ ID NO: 231, a vhCDR3 having SEQ ID NO: 232, a vlCDR1 having SEQ ID NO: 234, a vlCDR2 having SEQ ID NO: 235, and a vlCDR3 having SEQ ID NO: 236; and
iii) a vhCDR1 having SEQ ID NO: 238, a vhCDR2 having SEQ ID NO: 239, a vhCDR3 having SEQ ID NO: 240, a vlCDR1 having SEQ ID NO: 242, a vlCDR2 having SEQ ID NO: 243, and a vlCDR3 having SEQ ID NO: 244.
2. The composition of claim 1 , wherein the variable heavy domain has SEQ ID NO: 245 and the variable light domain has SEQ ID NO: 249.
3. The composition of claim 1 , wherein the variable heavy domain has SEQ ID NO: 229 and the variable light domain has SEQ ID NO: 233.
4. The composition of claim 1 , wherein the variable heavy domain has SEQ ID NO: 237 and the variable light domain has SEQ ID NO: 241.
5. A nucleic acid composition comprising:
a) a first nucleic acid encoding a variable heavy domain; and
b) a second nucleic acid encoding a variable light domain,
wherein the variable heavy domain and variable light domain are as set forth in any one of claims 2 - 4 .
6. An expression vector composition comprising:
a) a first expression vector comprising a nucleic acid encoding a variable heavy domain; and
b) a second expression vector comprising a nucleic acid encoding a variable light domain,
wherein the variable heavy domain and variable light domain are as set forth in any one of claims 2 - 4 .
7. A nucleic acid composition comprising:
a) a first nucleic acid encoding a variable heavy domain; and
b) a second nucleic acid encoding a variable light domain,
wherein the variable heavy domain vhCDR1-3 and the variable light domain v1CDR1-3 are selected from part b)i), part b)ii), or part b)iii) of claim 1 .
8. An expression vector composition comprising:
a) a first expression vector comprising a first nucleic acid encoding a variable heavy domain; and
b) a second expression vector comprising second nucleic acid encoding a variable light domain,
wherein the variable heavy domain vhCDR1-3 and the variable light domain v1CDR1-3 are selected from part b)i), part b)ii), or part b)iii).
9. A host cell comprising the expression vector composition of claim 8 .
10. A method of making a FAP binding domain comprising:
culturing the host cell of claim 9 under conditions wherein the first variable heavy domain and first variable light domain are expressed and the FAP binding domain is formed; and
purifying the FAP binding domain.
11. A heterodimeric antibody comprising:
a) a first monomer comprising, from N- to C-terminus, a VH1-CH1-linker 1-scFv-linker 2-CH2-CH3, wherein CH2-CH3 is a first Fc domain, and wherein linker 1 and linker 2 are a first linker and a second linker, respectively;
b) a second monomer comprising, from N- to C-terminus, a VH1-CH1-hinge-CH2-CH3, wherein CH2-CH3 is a second Fc domain; and
c) a common light chain comprising, from N- to C-terminus, a VL1-CL,
wherein VH1 is a first variable heavy domain and VL1 is a first variable light domain and VH1 and VL1 form two FAP binding domains;
wherein the first variable heavy domain and first variable light domain are selected from the following:
i) a first variable heavy domain having SEQ ID NO: 245 and a first variable light domain having SEQ ID NO: 249;
ii) a first variable heavy domain having SEQ ID NO: 229 and a first variable light domain having SEQ ID NO: 233; and
iii) a first variable heavy domain having SEQ ID NO: 237 and a first variable light domain having SEQ ID NO: 241,
wherein the scFv is an anti-CD3 scFv comprising a second variable heavy domain (VH2) covalently attached to a second variable light domain (VL2) by an scFv linker.
12. The heterodimeric antibody of claim 11 , wherein the scFv is in the orientation, from N- to C-terminus, VH2-scFv linker-VL2.
13. The heterodimeric antibody of claim 11 , wherein the scFv is in the orientation, from N- to C-terminus, VL2-scFv linker-VH2.
14. The heterodimeric antibody of claim 11 , wherein the second variable heavy domain and second variable light domain are selected from the following:
a) a second variable heavy domain having SEQ ID NO: 73 and a second variable light domain having SEQ ID NO: 79;
b) a second variable heavy domain having SEQ ID NO: 85 and a second variable light domain having SEQ ID NO: 89;
c) a second variable heavy domain having SEQ ID NO: 95 and a second variable light domain having SEQ ID NO: 99;
d) a second variable heavy domain having SEQ ID NO: 105 and a second variable light domain having SEQ ID NO: 109;
e) a second variable heavy domain having SEQ ID NO: 115 and a second variable light domain having SEQ ID NO: 119; and
f) a second variable heavy domain having SEQ ID NO: 125 and a second variable light domain having SEQ ID NO: 129.
15. The heterodimeric antibody of claim 11 , wherein the first Fc domain is a first variant human IgG Fc domain and the second Fc domain is a second variant human IgG Fc domain.
16. The heterodimeric antibody of claim 15 , wherein the first variant human IgG Fc domain comprises amino acid substitutions S364K/E357Q and the second variant human IgG Fc domain comprises amino acid substitutions L368D/K370S, wherein numbering is according to EU numbering.
17. The heterodimeric antibody of claim 15 , wherein the first variant human IgG Fc domain and the second variant human IgG Fc domain each comprise amino acid substitutions E233P/L234V/L235A/G236del/S267K, wherein numbering is according to EU numbering.
18. The heterodimeric antibody of claim 15 , wherein the first variant human IgG Fc domain comprises amino acid substitutions S364K/E357Q and the second variant human IgG Fc domain comprises amino acid substitutions L368D/K370S,
wherein the first variant human IgG Fc domain and the second variant human IgG Fc domain each further comprise amino acid substitutions E233P/L234V/L235A/G236de1/S267K,
wherein the CH1-hinge-CH2-CH3 of the second monomer comprises amino acid substitutions N208D/Q295E/N384D/Q418E/N421D, wherein numbering is according to EU numbering.
19. The heterodimeric antibody of claim 18 , wherein the first variant human IgG Fc domain and the second variant human IgG Fc domain each further comprise amino acid substitutions M428L/N434S, wherein numbering is according to EU numbering.
20. A nucleic acid composition comprising:
a) a first nucleic acid encoding a first monomer;
b) a second nucleic acid encoding a second monomer; and
c) a third nucleic acid encoding a common light chain,
wherein the first monomer comprises, from N- to C-terminus, a VH1-CH1-linker 1-scFv-linker 2-CH2-CH3, wherein CH2-CH3 is a first Fc domain, and wherein linker 1 and linker 2 are a first linker and a second linker, respectively,
wherein the second monomer comprises, from N- to C-terminus, a VH1-CH1-hinge-CH2-CH3, wherein CH2-CH3 is a second Fc domain,
wherein the common light chain comprises, from N- to C-terminus, a VL1-CL,
wherein VH1 is a first variable heavy domain and VL1 is a first variable light domain and VH1 and VL1 form two FAP binding domains;
wherein the first variable heavy domain and first variable light domain are selected from the following:
i) a first variable heavy domain having SEQ ID NO: 245 and a first variable light domain having SEQ ID NO: 249;
ii) a first variable heavy domain having SEQ ID NO: 229 and a first variable light domain having SEQ ID NO: 233; and
iii) a first variable heavy domain having SEQ ID NO: 237 and a first variable light domain having SEQ ID NO: 241, and
wherein the scFv is an anti-CD3 scFv comprising a second variable heavy domain (VH2) covalently attached to a second variable light domain (VL2) by an scFv linker.
21. An expression vector composition comprising:
a) a first expression vector comprising a first nucleic acid encoding a first monomer;
b) a second expression vector comprising a second nucleic acid encoding a second monomer; and
c) a third expression vector comprising a third nucleic acid encoding a common light chain,
wherein the first monomer comprises, from N- to C-terminus, a VH1-CH1-linker 1-scFv-linker 2-CH2-CH3, wherein CH2-CH3 is a first Fc domain, and wherein linker 1 and linker 2 are a first linker and a second linker, respectively,
wherein the second monomer comprises, from N- to C-terminus, a VH1-CH1-hinge-CH2-CH3, wherein CH2-CH3 is a second Fc domain,
wherein the common light chain comprises, from N- to C-terminus, a VL1-CL,
wherein VH1 is a first variable heavy domain and VL1 is a first variable light domain and VH1 and VL1 form two FAP binding domains;
wherein the first variable heavy domain and first variable light domain are selected from the following:
i) a first variable heavy domain having SEQ ID NO: 245 and a first variable light domain having SEQ ID NO: 249;
ii) a first variable heavy domain having SEQ ID NO: 229 and a first variable light domain having SEQ ID NO: 233; and
iii) a first variable heavy domain having SEQ ID NO: 237 and a first variable light domain having SEQ ID NO: 241, and
wherein the scFv is an anti-CD3 scFv comprising a second variable heavy domain (VH2) covalently attached to a second variable light domain (VL2) by an scFv linker.
22. A host cell comprising the expression vector composition of claim 21 .
23. A method of making a heterodimeric antibody comprising:
a) culturing the host cell of claim 22 under conditions wherein the first monomer, the second monomer and the common light chain are expressed and the heterodimeric antibody is formed; and
b) purifying the heterodimeric antibody.