T cell receptor-like antibody agents specific for EBV latent membrane protein 2A peptide presented by human HLA
Described herein are antibodies, fragments thereof and multi-specific binding agents that bind an Epstein-Barr virus (EBV) latent membrane protein 2 (LMP2) peptide presented by HLA class I molecules, in particular, HLA-A02. Also provided herein are methods of using the same or compositions thereof for the detection, prevention and/or therapeutic treatment of diseases characterized by expression of an EBV-LMP2 peptide presented by HLA-A02, in particular, Burkit's lymphoma, Hodgkin's lymphoma and nasopharyngeal carcinoma.
1. A human antibody agent that binds an EBV-LMP2/HLA peptide complex comprising
(a) heavy chain CDR1 of SEQ ID NO:31, heavy chain CDR2 of SEQ ID NO:33 and heavy chain CDR3 of SEQ ID NO:35; and light chain CDR1 of SEQ ID NO:73, light chain CDR2 of SEQ ID NO:75 and light chain CDR3 of SEQ ID NO:77;
(b) heavy chain CDR1 of SEQ ID NO:37, heavy chain CDR2 of SEQ ID NO:39 and heavy chain CDR3 of SEQ ID NO:41; and light chain CDR1 of SEQ ID NO:79, light chain CDR2 of SEQ ID NO:81 and light chain CDR3 of SEQ ID NO:83;
(c) heavy chain CDR1 of SEQ ID NO:43, heavy chain CDR2 of SEQ ID NO:45 and heavy chain CDR3 of SEQ ID NO:47; and light chain CDR1 of SEQ ID NO:85, light chain CDR2 of SEQ ID NO:87 and light chain CDR3 of SEQ ID NO:89;
(d) heavy chain CDR1 of SEQ ID NO:49, heavy chain CDR2 of SEQ ID NO:51 and heavy chain CDR3 of SEQ ID NO:53; and light chain CDR1 of SEQ ID NO:91, light chain CDR2 of SEQ ID NO:93 and light chain CDR3 of SEQ ID NO:95;
(e) heavy chain CDR1 of SEQ ID NO:55, heavy chain CDR2 of SEQ ID NO:57 and heavy chain CDR3 of SEQ ID NO:59; and light chain CDR1 of SEQ ID NO:97, light chain CDR2 of SEQ ID NO:99 and light chain CDR3 of SEQ ID NO:101;
(f) heavy chain CDR1 of SEQ ID NO:61, heavy chain CDR2 of SEQ ID NO:63 and heavy chain CDR3 of SEQ ID NO:65; and light chain CDR1 of SEQ ID NO:103, light chain CDR2 of SEQ ID NO:105 and light chain CDR3 of SEQ ID NO:107; or
(g) heavy chain CDR1 of SEQ ID NO:67, heavy chain CDR2 of SEQ ID NO:69 and heavy chain CDR3 of SEQ ID NO:71; and light chain CDR1 of SEQ ID NO:109, light chain CDR2 of SEQ ID NO:111 and light chain CDR3 of SEQ ID NO:113.
2. The human antibody agent of claim 1 , wherein the human antibody agent has a K D of about 2.0 to about 170 nM.
3. The human antibody agent of claim 1 , wherein the EBV-LMP2 peptide has an amino acid sequence that is or comprises CLGGLLTMV (SEQ ID NO:1).
4. The human antibody agent of claim 3 , wherein the human antibody agent interacts directly with amino acid residues at one or more positions selected from the group consisting of position 1, 2, 3, 4, 5, 6, 7, 8, and combinations thereof, in the EBV-LMP2 peptide.
5. The human antibody agent of claim 1 , wherein the human antibody agent interacts directly with amino acid residues at one or more positions selected from the group consisting of position 1, position 5, position 8, and combinations thereof, in the EBV-LMP2 peptide.
6. The human antibody agent of claim 1 , wherein the human antibody agent comprises
(a) a heavy chain variable region sequence as set forth in any one of SEQ ID NOs: 5, 9, 13, 17, 21, 25, and 29, and
(b) a light chain variable region sequence as set forth in any one of SEQ ID NOs: 3, 7, 11, 15, 19, 23, and 27.
7. The human antibody agent of claim 1 , wherein the human antibody agent comprises
(a) the light chain variable region of SEQ ID NO:3 and the heavy chain variable region of SEQ ID NO:5;
(b) the light chain variable region of SEQ ID NO:7 and the heavy chain variable region of SEQ ID NO:9;
(c) the light chain variable region of SEQ ID NO:11 and the heavy chain variable region of SEQ ID NO:13;
(d) the light chain variable region of SEQ ID NO:15 and the heavy chain variable region of SEQ ID NO:17;
(e) the light chain variable region of SEQ ID NO:19 and the heavy chain variable region of SEQ ID NO:21;
(f) the light chain variable region of SEQ ID NO:23 and the heavy chain variable region of SEQ ID NO:25; or
(g) the light chain variable region of SEQ ID NO:27 and the heavy chain variable region of SEQ ID NO:29.
8. The human antibody agent of claim 1 , wherein the human antibody agent is a human monoclonal antibody that is defucosylated and/or glycosylated with terminal mannose, N-acetylglucose or glucose.
9. The human antibody agent of claim 1 , which human antibody agent comprises one or more amino acid substitutions that increase affinity to the EBV-LMP2/HLA peptide complex, wherein the one or more amino acid substitutions is selected from a light chain I48V substitution, a light chain K66R substitution, a heavy chain V5E substitution, a heavy chain E10D substitution, a heavy chain G26E substitution, and a heavy chain V78A substitution.
10. The human antibody agent of claim 9 , which human antibody agent that comprises one or more amino acid substitutions is characterized by
(a) a KA of at least about 1.6 higher affinity than affinity to a reference peptide/HLA complex, and/or
(b) no cross-reactivity to a reference peptide/HLA complex.
11. The human antibody agent of claim 1 , wherein the human antibody agent is a human monoclonal antibody or a fragment thereof.
12. The human antibody agent of claim 11 , wherein the human monoclonal antibody is an IgG1.
13. The human antibody agent of claim 11 , wherein the human monoclonal antibody fragment is an scFv.
14. A bispecific antibody comprising a first antigen-binding site that binds an EBV-LMP2/HLA peptide complex and a second antigen-binding site, wherein the first antigen-binding site comprises
(a) heavy chain CDR1 of SEQ ID NO:31, heavy chain CDR2 of SEQ ID NO:33 and heavy chain CDR3 of SEQ ID NO:35; and light chain CDR1 of SEQ ID NO:73, light chain CDR2 of SEQ ID NO:75 and light chain CDR3 of SEQ ID NO:77;
(b) heavy chain CDR1 of SEQ ID NO:37, heavy chain CDR2 of SEQ ID NO:39 and heavy chain CDR3 of SEQ ID NO:41; and light chain CDR1 of SEQ ID NO:79, light chain CDR2 of SEQ ID NO:81 and light chain CDR3 of SEQ ID NO:83;
(c) heavy chain CDR1 of SEQ ID NO:43, heavy chain CDR2 of SEQ ID NO:45 and heavy chain CDR3 of SEQ ID NO:47; and light chain CDR1 of SEQ ID NO:85, light chain CDR2 of SEQ ID NO:87 and light chain CDR3 of SEQ ID NO:89;
(d) heavy chain CDR1 of SEQ ID NO:49, heavy chain CDR2 of SEQ ID NO:51 and heavy chain CDR3 of SEQ ID NO:53; and light chain CDR1 of SEQ ID NO:91, light chain CDR2 of SEQ ID NO:93 and light chain CDR3 of SEQ ID NO:95;
(e) heavy chain CDR1 of SEQ ID NO:55, heavy chain CDR2 of SEQ ID NO:57 and heavy chain CDR3 of SEQ ID NO:59; and light chain CDR1 of SEQ ID NO:97, light chain CDR2 of SEQ ID NO:99 and light chain CDR3 of SEQ ID NO:101;
(f) heavy chain CDR1 of SEQ ID NO:61, heavy chain CDR2 of SEQ ID NO:63 and heavy chain CDR3 of SEQ ID NO:65; and light chain CDR1 of SEQ ID NO:103, light chain CDR2 of SEQ ID NO:105 and light chain CDR3 of SEQ ID NO:107; or
(g) heavy chain CDR1 of SEQ ID NO:67, heavy chain CDR2 of SEQ ID NO:69 and heavy chain CDR3 of SEQ ID NO:71, and light chain CDR1 of SEQ ID NO:109, light chain CDR2 of SEQ ID NO:111 and light chain CDR3 of SEQ ID NO:113.
15. The bispecific antibody of claim 14 , wherein the bispecific antibody is characterized by an EC 50 of about 0.2 to about 135 pM in T2 cells pulsed with EBV-LMP2 peptide CLGGLLTMV (SEQ ID NO:1).
16. The bispecific antibody of claim 14 , wherein the first and/or second antigen-binding sites are selected from the group consisting of an immunoglobulin molecule, scFv, scFab, Fab, Fv or a combination thereof.
17. The bispecific antibody of claim 14 , wherein the first and second antigen-binding sites are configured such that they form a single polypeptide chain.
18. The bispecific antibody of claim 17 , wherein the first and second antigen-binding sites are each scFvs.
19. The bispecific antibody of claim 14 , wherein the second antigen-binding site binds an immunological cell selected from the group consisting of a T cell, B cell, NK cell, dendritic cell, monocyte, macrophage, neutrophil, mesenchymal stem cell and neural stem cell.
20. The bispecific antibody of claim 19 , wherein the second antigen-binding site binds CD3 on T cells.
21. The bispecific antibody of claim 14 , comprising
a first scFv that binds to EBV-LMP2 peptide CLGGLLTMV (SEQ ID NO:1) presented by an HLA-A02 molecule, and
a second scFv that binds CD3 on T cells,
wherein the bispecific antibody is characterized by an EC 50 of about 0.2 to about 135 pM in T2 cells pulsed with the EBV-LMP2 peptide.
22. The bispecific antibody of claim 21 , wherein the first scFv comprises
(a) the light chain variable region of SEQ ID NO:3 and the heavy chain variable region of SEQ ID NO:5;
(b) the light chain variable region of SEQ ID NO:7 and the heavy chain variable region of SEQ ID NO:9;
(c) the light chain variable region of SEQ ID NO:11 and the heavy chain variable region of SEQ ID NO:13;
(d) the light chain variable region of SEQ ID NO:15 and the heavy chain variable region of SEQ ID NO:17;
(e) the light chain variable region of SEQ ID NO:19 and the heavy chain variable region of SEQ ID NO:21;
(f) the light chain variable region of SEQ ID NO:23 and the heavy chain variable region of SEQ ID NO:25; or
(g) the light chain variable region of SEQ ID NO:27 and the heavy chain variable region of SEQ ID NO:29.
23. The bispecific antibody of claim 21 , wherein the HLA-A02 molecule is selected from HLA-A*02:01, HLA-A*02:02, HLA-A*02:03, HLA-A*02:04, HLA-A*02:05 and HLA-A*02:06.
24. The bispecific antibody of claim 21 , wherein the first scFv of the bispecific antibody interacts directly with at least position 5 of the EBV-LMP2 peptide.
25. A human antibody agent that binds an EBV-LMP2/HLA peptide complex comprising
(i) heavy chain CDR1 of SEQ ID NO:43, heavy chain CDR2 of SEQ ID NO:45 or a sequence that differs from SEQ ID NO:45 in an I51V substitution, and heavy chain CDR3 of SEQ ID NO:47; and
(ii) light chain CDR1 of SEQ ID NO:85, light chain CDR2 of SEQ ID NO:87 or a sequence that differs from SEQ ID NO:87 in one or both of a S52G and P55H substitution, and light chain CDR3 of SEQ ID NO:89 or a sequence that differs from SEQ ID NO:89 in a N95I substitution.
26. The human antibody agent of claim 25 , comprising
(i) a light chain variable region sequence that comprises a sequence of SEQ ID NO: 11 or a sequence that differs from SEQ ID NO:11 in one or more amino acid substitutions selected from I48V, S52G, P55H, K66R and N95I; and
(ii) a heavy chain variable region that comprises a sequence of SEQ ID NO: 13 or a sequence that differs from SEQ ID NO:13 in one or more amino acid substitutions selected from V5E, E10D, G26E, I51V and V78A.
27. A bispecific antibody comprising a first antigen-binding site that binds an EBV-LMP2/HLA peptide complex and a second antigen-binding site, wherein the first antigen-binding site comprises
(i) heavy chain CDR1 of SEQ ID NO:43, heavy chain CDR2 of SEQ ID NO:45 or a sequence that differs from SEQ ID NO:45 in an I51V substitution, and heavy chain CDR3 of SEQ ID NO:47; and
(ii) light chain CDR1 of SEQ ID NO:85, light chain CDR2 of SEQ ID NO:87 or a sequence that differs from SEQ ID NO:87 in one or both of a S52G and P55H substitution, and light chain CDR3 of SEQ ID NO:89 or a sequence that differs from SEQ ID NO:89 in a N95I substitution.
28. The bispecific antibody of claim 27 , wherein the first antigen-binding site comprises
(i) a light chain variable region sequence that comprises a sequence of SEQ ID NO: 11 or a sequence that differs from SEQ ID NO:11 in one or more amino acid substitutions selected from I48V, S52G, P55H, K66R and N95I; and
(ii) a heavy chain variable region that comprises a sequence of SEQ ID NO: 13 or a sequence that differs from SEQ ID NO:13 in one or more amino acid substitutions selected from V5E, E10D, G26E, I51V and V78A.