TGFβ antibodies, methods and uses
ProTGFβ1-GARP complex-selective antibodies, polynucleotides capable of encoding the proTGFβ1-GARP complex-selective antibodies or antigen-binding fragments, cells expressing proTGFβ1-GARP complex-selective antibodies or antigen-binding fragments, as well as associated vectors and detectably labeled proTGFβ1-GARP complex-selective antibodies or antigen-binding fragments may be used to enhance an immune response in a subject, for example, against a cancer.
1. An antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof specifically binds to human proTGFβ1 in a complex with human glycoprotein A repetitions predominant (proTGFβ1-GARP complex), wherein the antibody or antigen-binding fragment thereof comprises:
a. a heavy chain complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 6, and a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 7, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 9; or
b. a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, and a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 13, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15.
2. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof inhibits regulatory T cell (Treg) function in vitro.
3. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof inhibits the activation of TGFβ1.
4. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof binds to an epitope of human proTGFβ1 modified as a result of complex formation with human GARP.
5. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof binds to a polypeptide comprising the amino acid sequence of SEQ ID NO: 2 in the presence of a polypeptide comprising the amino acid sequence of SEQ ID NO: 1.
6. The antibody or antigen-binding fragment thereof of claim 5 , wherein the antibody or antigen-binding fragment thereof specifically binds to human proTGFβ1 with a binding affinity of at least 880 pM as measured by biolayer interferometry assay.
7. The antibody or antigen-binding fragment thereof of claim 5 , wherein the antibody or antigen-binding fragment thereof binds to human proTGFβ1 with a dissociation constant (Kd) of less than or equal to 1 nM for human proTGFβ1 in a complex with human glycoprotein A repetitions predominant (proTGFβ1-GARP complex) and wherein said proTGFβ1-GARP complex is in solution.
8. The antigen-binding fragment thereof of claim 1 , wherein the antigen-binding fragment is a Fab fragment, a Fab2 fragment, or a single chain antibody.
9. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment is recombinant.
10. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof are of lgG1, lgG2, lgG3, or lgG4 isotype.
11. The antibody or antigen-binding fragment thereof of claim 10 , wherein the antibody is an lgG4 isotype.
12. A polynucleotide encoding the antibody or antigen-binding fragment thereof of claim 1 .
13. A vector comprising the polynucleotide of claim 12 .
14. A host cell comprising the vector of claim 13 .
15. A process for the production of an antibody or antigen-binding fragment thereof, comprising:
culturing the host cell of claim 14 under the conditions allowing the expression of the antibody or antigen-binding fragment thereof, and
recovering the antibody or antigen-binding fragment thereof from the culture.
16. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.
17. A kit comprising the antibody or antigen-binding fragment thereof of claim 1 and packaging for the same.
18. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment specifically binds to human proTGFβ1 in a complex with human glycoprotein A repetitions predominant (proTGFβ1-GARP complex);
wherein said complex is in solution;
wherein the antibody or antigen-binding fragment thereof has no detectable binding according to a biolayer interferometry assay to any of the following:
a TGFβ1 growth factor domain,
a TGFβ2 growth factor domain,
a TGFβ3 growth factor domain,
a proTGFβ1 covalently associated with LTBP1,
a proTGFβ1 covalently associated with LTBP3,
a proTGFβ1 covalently associated with LRRC33, or
a proTGFβ1 that is not associated with human GARP; and
wherein the antibody or antigen-binding fragment thereof has an inhibitory concentration (IC50) of less than or equal to 10 nM for inhibition of TGFB1 growth factor release from a cell-associated proTGFβ1-GARP complex.
19. A method of treating a subject with an infectious disease or hyperproliferative disorder comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof of claim 1 .
20. The method of claim 19 , wherein the antibody or antigen-binding fragment thereof is administered in combination with one or more additional therapies or therapeutic agents selected from:
(a) a chemotherapeutic agent,
(b) radiotherapy, and
(c) a T-cell checkpoint inhibitor selected from an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.
21. The method of claim 19 , wherein the hyperproliferative disorder is cancer.
22. The method of claim 21 , wherein the cancer is kidney cancer or renal cell carcinoma.
23. The method of claim 21 , wherein the treatment results in one or more of the following:
inhibition of further tumor growth,
induction of tumor regression,
increase of progression-free survival,
extension of overall survival, or
delay or prevention of the onset of metastasis.
24. An antibody or antigen-binding fragment thereof, comprising:
a. a heavy chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 17,
wherein the antibody or antigen-binding fragment thereof comprises heavy chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 4-6, respectively, and light chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 7-9, respectively; or
b. a heavy chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 18 and a light chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 19,
wherein the antibody or antigen-binding fragment thereof comprises heavy chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 10-12, respectively, and light chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 13-15, respectively.
25. The antibody or antigen-binding fragment thereof of claim 24 , wherein
a. the heavy chain comprises the amino acid sequence of SEQ ID NO: 16 and the light chain comprises the amino acid sequence of SEQ ID NO: 17; or
b. the heavy chain comprises the amino acid sequence of SEQ ID NO: 18 and the light chain comprises the amino acid sequence of SEQ ID NO: 19.
26. An antibody or antigen-binding fragment thereof, comprising:
a. a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to amino acids 1-118 of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90% identity to amino acids 1-107 of SEQ ID NO: 17,
wherein the antibody or antigen-binding fragment thereof comprises heavy chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively, and light chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively;
b. a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to amino acids 1-121 of SEQ ID NO: 18 and a light chain variable region comprising an amino acid sequence that is at least 90% identical to amino acids 1-110 of SEQ ID NO: 19,
wherein the antibody or antigen-binding fragment thereof comprises heavy chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 10-12, respectively, and light chain CDRs 1-3 comprising the amino acid sequences of SEQ ID NOs: 13-15, respectively;
c. a heavy chain variable region comprising amino acids 1-118 of SEQ ID NO: 16 and a light chain variable region sequence comprising amino acids 1-107 of SEQ ID NO: 17; or
d. a heavy chain variable region comprising amino acids 1-121 of SEQ ID NO: 18 and a light chain variable region sequence comprising amino acids 1-110 of SEQ ID NO: 19.