Antibody against HLA-g

Abstract

The present invention relates to antibodies, or antigen-binding fragments thereof, that are capable of binding to human leukocyte antigen G (HLA-G). The present invention also relates to methods of producing and therapeutic uses of such antibodies, or antigen-binding fragments thereof.

Description

[0001] ANTIBODY AGAINST HLA-G

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to antibodies, or antigen-binding fragments thereof, that are capable of binding to human leukocyte antigen G (HLA-G). The present invention also relates to methods of producing and therapeutic uses of such antibodies, or antigen-binding fragments thereof.

[0004] BACKGROUND TO THE INVENTION

[0005] Antibodies are glycoproteins belonging to the immunoglobulin superfamily of proteins. There are five primary classes of immunoglobulin, including IgG, IgM, IgA, I g E and IgD. Antibodies are typically made of basic structural units comprising two heavy and light chains. The IgG immunoglobulin molecule, for example, comprises four polypeptide chains, two of which are identical heavy (H) chains and two of which are identical light (K) chains. Each heavy chain comprises an N-terminal variable domain (VH) and three constant domains (CH1 , CH2 and CH3), with a hinge region linking CH1 and CH2. Each light chain comprises an N-terminal variable domain (VL) and a constant domain (CL), which together associate with the VH and CH1 domain of the heavy chain.

[0006] An antibody may recognise an antigen via the fragment antigen-binding (Fab) variable region, which comprises the VL, CL, VH and CH1 domains. The antibody may activate the immune system through the fragment crystallisable region (Fc region), which is the tail region of the antibody comprising the CH2 and CH3 domains.

[0007] The heavy chain variable region (VH) refers to the fragment of the heavy chain of an antibody that comprises the three complementarity determining regions (CDRs) which are interposed between the framework regions, the latter of which form a scaffold which supports the CDRs. Similarly, the light chain variable region (VL) comprises three CDRs and their surrounding framework regions.

[0008] The CDR, with regard to an antibody or an antigen-binding fragment thereof, is a highly variable sequence of amino acids that form a loop in the variable regions of the heavy and light chains. The CDR is therefore able to interact with the antigen. The heavy and light chain variable regions each contain three CDRs, that are referred to herein as CDRH1 , CDRH2 and CDRH3 (heavy chain CDRs) and CDRL1 , CDRL2 and CDRL3 (light chain CDRs).

[0009] Human leukocyte antigen-G (HLA-G) is a non-classical Major Histocompatibility Complex class I (MHCI). Classical MHCI molecules are responsible for the presentation of antigenic peptide ligands on infected cells to CD8 positive T cells. In contrast, a key role of non-Classical MHCI is to mediate inhibitory or activating stimuli to Natural Killer (NK) cells. The major classical MHC molecules are HLA-A and HLA-B, whereas the major non-classical MHC molecules are HLA-C, HLA-E, HLA-F and HLA-G. HLA-G is expressed on placental trophoblasts and thymic epithelial cells and is therefore responsible for the suppression of a wide-range of immune responses at the maternal-fetal interface. HLA-G, HLA-E and HLA-C suppress maternal immune responses through binding leukocyte Ig-like receptors (LILRs), such as LILRB2, which is expressed on, for example, monocytes, macrophages and granulocytes (HoWangYin KY et al. Cell Mol Life Sci. 2012 Dec;69(23):4041-9).

[0010] LI LRB2 contains four tandem Ig-like domains and is a receptor for classical and non-classical M HC- I molecules, such as HLA-G. LILRB2 is part of the LILR family, of which there are 11 members (LILRB1, LILRB2, LILRB3, LILRB4, LILRB5, LILRA1 , LILRA2, LILRA3, LILRA4, LILRA5 and LILRA6). LILRB1 to LILRB5 are referred to as inhibitory receptors, whereas LILRA1 to LILRA6 (excluding A3) are referred to as activating receptors. LILRB2 has immunoreceptor tyrosine-based inhibitory motifs in its cytoplasmic tail, which enables recruitment of the protein tyrosine phosphatase SHP-1 to result in inhibitory signalling.

[0011] Although the HLA-G - LILRB2 interaction is responsible for initiating immunosuppressive pathways that shield the developing fetus from the mother’s immune system, HLA-G has also been shown to be expressed as a tumour specific antigen (Loustau M et al. Front Immunol. 2020 Aug 14;11 :1685). HLA-G may therefore be expressed on the surface of tumour cells, whereupon binding to LILRB2 on immune cells induces an immunosuppressive effect that enables immune escape of the cancerous cells.

[0012] Therefore, there remains a need for molecules that prevent the interaction between HLA-G and LILRB2.

[0013] SUMMARY OF THE INVENTION

[0014] The present invention is based, at least in part, on the inventors’ development of antibodies which have particularly advantageous and surprising properties.

[0015] It is desirable to produce antibodies that comprise HLA-G binding domains that bind specifically to HLA-G and not the other 5 class I MHC complexes (HLA-A, -B, -C, -E and -F).

[0016] The present inventors have developed an antibody, or antigen-binding fragment thereof, that targets HLA-G and may be used as an immunogenic agent or used in the manufacture of an immunogenic agent that may be capable of treating cancer. The present inventors have surprisingly demonstrated that the antibodies, or antigen-binding fragments thereof, of the present invention are capable of preventing the immunosuppressive effect of HLA-G expression on the surface of tumour cells.

[0017] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising:

[0018] (a) a CDRH1 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 37;

[0019] (b) a CDRH2 selected from the group consisting of SEQ ID NO 38 to SEQ ID NO 77; and

[0020] (c) a CDRH3 selected from the group consisting of SEQ ID NO 78 to SEQ ID NO 127; and a light chain variable region comprising:

[0021] (d) a CDRL1 selected from the group consisting of SEQ ID NO: 128 to SEQ ID NO: 168;

[0022] (e) a CDRL2 selected from the group consisting of SEQ ID NO: 169 to SEQ ID NO: 192; and

[0023] (f) a CDRL3 selected from the group consisting of SEQ ID NO: 193 to SEQ ID NO: 238.

[0024] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprise CDRs selected from one of the following:

[0025] (a) CDRHs according to SEQ ID NO: 1, 38 and 78;

[0026] (b) CDRHs according to SEQ ID NO: 2, 39 and 79;

[0027] (c) CDRHs according to SEQ ID NO: 3, 40 and 80;

[0028] (d) CDRHs according to SEQ ID NO: 9, 74 and 123;

[0029] (e) CDRHs according to SEQ ID NO: 6, 43 and 83;

[0030] (f) CDRHs according to SEQ ID NO: 16, 52 and 93;

[0031] (g) CDRHs according to SEQ ID NO: 19, 54 and 96;

[0032] (h) CDRHs according to SEQ ID NO: 8, 44 and 85;

[0033] (i) CDRHs according to SEQ ID NO: 12, 48 and 89;

[0034] (j) CDRHs according to SEQ ID NO: 13, 64 and 107;

[0035] (k) CDRHs according to SEQ ID NO: 33, 54 and 118;

[0036] (l) CDRHs according to SEQ ID NO: 13, 40 and 121 ;

[0037] (m) CDRHs according to SEQ ID NO: 13, 76 and 125; or

[0038] (n) CDRHs according to SEQ ID NO: 37, 40 and 126. In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the light chain variable region comprises CDRs selected from one of the following:

[0039] (a) CDRLs according to SEQ ID NO: 128 , 169 and 193;

[0040] (b) CDRLs according to SEQ ID NO: 129 , 170 and 194;

[0041] (c) CDRLs according to SEQ ID NO: 130 , 171 and 195;

[0042] (d) CDRLs according to SEQ ID NO: 143 , 190 and 234;

[0043] (e) CDRLs according to SEQ ID NO: 133 , 178 and 198;

[0044] (f) CDRLs according to SEQ ID NO: 142 , 171 and 207;

[0045] (g) CDRLs according to SEQ ID NO: 145 , 182 and 210;

[0046] (h) CDRLs according to SEQ ID NO: 135 , 175 and 200;

[0047] (i) CDRLs according to SEQ ID NO: 139 , 179 and 203;

[0048] (j) CDRLs according to SEQ ID NO: 154 , 169 and 220;

[0049] (k) CDRLs according to SEQ ID NO: 143 , 181 and 229;

[0050] (l) CDRLs according to SEQ ID NO: 165 , 171 and 232;

[0051] (m) CDRLs according to SEQ ID NO: 143 , 171 and 236; or

[0052] (n) CDRLs according to SEQ ID NO: 162 , 191 and 237.

[0053] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 1 , 38 and

[0054] 78 and CDRLs according to SEQ ID NO: 128, 169 and 193.

[0055] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 239, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 289.

[0056] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 2, 39 and

[0057] 79 and CDRLs according to SEQ ID NO: 129, 170 and 194.

[0058] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 240, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 290. In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 3, 40 and 80 and CDRLs according to SEQ ID NO: 130, 171 and 195.

[0059] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 241 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 291.

[0060] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 9, 74 and 123 and CDRLs according to SEQ ID NO: 143, 190 and 234.

[0061] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 285, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 335.

[0062] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 6, 43 and 83 and CDRLs according to SEQ ID NO: 133, 178 and 198.

[0063] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 247, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 297.

[0064] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 16, 52 and 93 and CDRLs according to SEQ ID NO: 142, 171 and 207.

[0065] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 256, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 306. In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 19, 54 and 96 and CDRLs according to SEQ ID NO: 145, 182 and 210.

[0066] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 259, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 309.

[0067] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 8, 44 and 85 and CDRLs according to SEQ ID NO: 135, 175 and 200.

[0068] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 249, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 299.

[0069] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 12, 48 and 89 and CDRLs according to SEQ ID NO: 139, 179 and 203.

[0070] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 252, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 302.

[0071] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 64 and 107 and CDRLs according to SEQ ID NO: 154, 169 and 220.

[0072] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 269, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 319. In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 33, 54 and 118 and CDRLs according to SEQ ID NO: 143, 181 and 229.

[0073] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 280, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 330.

[0074] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 40 and 121 and CDRLs according to SEQ ID NO: 165, 171 and 232.

[0075] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 283, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 333.

[0076] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 76 and 125 and CDRLs according to SEQ ID NO: 143, 171 and 236.

[0077] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 286, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 336.

[0078] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the variable region comprises CDRHs according to SEQ ID NO: 37, 40 and 126 and CDRLs according to SEQ ID NO: 162, 191 and 237.

[0079] In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 287, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 337. In one aspect, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, for use in the treatment or prevention of cancer.

[0080] BRIEF DESCRIPTION OF THE FIGURES

[0081] Figure 1 : On-target flow cytometric analysis of mAb binding to HLA-G-expressing HEK293T cells. Cells transfected with HLA-G were stained with anti-HLA-G mAb clones (thick line, open histogram) or human lgG1 isotype control (grey shaded histogram), followed by anti-human IgG- APC secondary antibody. Confirmation of HLA-G expression by HEK293T was demonstrated using an APC-conjugated commercial anti-HLA-G mAb.

[0082] Figure 2: Off-target flow cytometric analysis of mAb binding to H LA -expressing HEK293T cells. Cells transfected with HLA-A, HLA-B, HLA-C, HLA-E, or HLA-F were stained with anti-HLA- G mAb clones or human lgG1 isotype control, followed by anti-human IgG-APC secondary antibody. Data is presented as MFI of anti-HLA-G stain relative to isotype control.

[0083] Figure 3: Direct ELISA. Data shows anti-HLAG mAbs bind to recombinant human HLAG complex tetramer. The lower the EC50 the better binding the anti-HLAG is to recombinant human HLAG complex tetramer.

[0084] Figure 4: Competitive ELISA. Data shows anti-HLAG antibodies that block the binding of recombinant HLAG to recombinant LILRB2-Fc. The higher the percent blocking the better the capacity of the mAb to block binding of the two recombinant antigens.

[0085] Figure 5: Expression. Data shows that all mAb clones demonstrate expression above 200 mg / kg

[0086] Figure 6: Purity. Data shows that all mAbs are purified to an acceptable purity.

[0087] Figure 7: Thermostability. Data shows each mAb has good thermostability representing a good chemistry, manufacturing and controls (CMC) attribute.

[0088] Figure 8: TNF-o Expression. Data shows that mAbs are able to restore TNF-a expression in myeloid cells. In the presence of JEG3 cells (tumorigenic cells expressing HLA-G) and LPS, U937 cells (myeloid cells) express less TNF-a due to the immunosuppressive effect of JEG-3 cells. In the presence of mAbs, TNF- a expression is restored. Figure 9: KB17A.5.3 Binding to Human HLA-G by ELISA. KB17A.5.3 binds to recombinant human HLA-G with strong affinity (16.7 ng / mL) comparable to TTX-080 produced in Nicotians benthamiana with an affinity of 9.7 ng / mL.

[0089] Figure 10: KB17A.5.3 Human vs Cynomolgus and Human vs Rhesus Macaque HLA-G Binding. KB17A.5.3 did not demonstrate binding affinity for either Cynomolgus (upper graph) or Rhesus macaque (lower graph) HLA-G but bound with high affinity to human HLA-G. Antibody TTX- 080 produced in Nicotians benthamiana was used as a positive binding control. Human, Cynomolgus, and Rhesus macaque HLA-G recombinant antigens have confirmed binding to the corresponding species LILRB2 per the vendor specification sheet.

[0090] Figure 11 : KB17A.5.3 Binding to Human HLA-A, HLA-B, HLA-C, HLA-E and HLA-G. KB17A.5.3 binds to HLA-G overexpressing HEK293T cells down to a concentration of 0.0015 pg / mL; at 1.5 pg / mL, KB17A.5.3 does not bind to HLA -A, -B, -C, or -E.

[0091] Figure 12: KB17A.5.3 Binding to HLA-G Expressing JEG-3 Cells. KB17A.5.3 binds to JEG-3 cells, an endogenous HLA-G expressing cell-line, with an ECso of 78.6 ng / mL. Graphs were plotted and ECso’s were calculated with GraphPad Prism 8.0 software.

[0092] Figure 13: Binding Confirmation of KB17A.5.3 by Surface Plasmon Resonance (SPR). KB17A.5.3 binds to HLA-G with high affinity and corroborates the ELISA and flow cytometry binding data.

[0093] Figure 14: LILRB2 / HLA-G Blocking ELISA. KB17A.5.3 exhibited blocking activity with an IC50 of 0.338 pg / mL confirming that the antibody is able to block binding association between recombinant HLA-G and LILRB2.

[0094] Figure 15: Fc Receptor Binding by ELISA. KB17A.5.3 binds strongly to human Fc receptors CD64 (ECso, 38.61 ± 17.8 ng / mL) and CD16a (EC50, 108.05 ± 91.9 ng / mL) relative to positive control (commercially-obtained Humira). Additionally, KB17A.5.3 binds to CD32b / c (EC50, 171.5 pg / mL) and CD32a (EC50, 6.5 pg / mL).

[0095] Figure 16: FcRn Binding by ELISA. KB17A.5.3 binds to neonatal Fc Receptor with an EC50 of 170.8 pg / mL. Figure 17: Prevention of HLA-G Induced Monocyte Suppression. KB17A.5.3 prevents HLA-G induced monocyte suppression of TNF-a cytokine release in a dose-dependent manner when compared to controls in four donors. # indicates > 80% TNF-a release was maintained in comparison to stimulation control.

[0096] Figure 18: Effector Function Activity Using Primary Cells. KB17A.5.3 elicited cytolysis of HLA- G expressing JEG-3 when in co-culture with primary human PBMC cells. KB17A.5.3 demonstrated ADCC activity with PBMC cells in multiple donors - Donor 1 : EC50, 0.052 pg / mL (upper graph) and Donor 2: EC50, 0.023 pg / mL (lower graph).

[0097] Figure 19: Effector Functional Activity Using Natural Killer (NK) Cells. KB17A.5.3 elicited robust cytotoxicity of JEG-3 cells when primary human NK cells were co-cultured with HLA-G-expressing JEG-3 cells. In contrast, a plant-made lgG1 isotype control did not demonstrate any killing above background.

[0098] Figure 20: Effector Functional Activity Using Jurkat Cells. KB17A.5.3 elicited ADCC reporter gene activity in the assay after incubation periods of 6 hours (EC50, 0.064 pg / mL) and 23 hours (EC50, 0.070 pg / mL).

[0099] Figure 21 : Mean change (A) in tumor volume by treatment group with SEM. The treatment (20 mg / kg) was performed BIW for 31 days (10 total doses). The study was terminated on Day 38. The mean (SEM) of two CD34+ HSC donors from each dosing group is plotted (n=6).

[0100] Figure 22: Mean whole tumor weight at Day 38. The mean (±SEM) tumor weights are graphed. P<0.05 is considered statistically significant compared with “Isotype group”.

[0101] Figure 23: Absolute Count and Percentage of T cell Subsets. Absolute count result (top) and percentage (bottom) of T cell subsets in tumor samples per flow cytometry analysis. P<0.05 is considered statistically significant compared with “Isotype group”.

[0102] Figure 24: Absolute Count and Percentage of Myeloid Cell Substrates. Absolute count result (top) and percentage (bottom) of myeloid cell subsets in tumor samples per flow cytometry analysis. P<0.05 is considered statistically significant compared with “Isotype group”.

[0103] Figure 25: HLA-G Expression in Tumors. Western Blot Assay Depicting HLA-G Expression in Tumors. Tumor samples were harvested, flash frozen, and stored at -80°C until analysis. HLA-G was detected using the anti-HLA-G antibody 4H84 (ab52455). Left, hlgG1 Isotype Control tumor samples labeled SK-MEL-5hHLA-G-#8-G1. Right, KB17A.5.3 tumor samples labeled SK-MEL- 5hHLA-G-#8-G2. JEG-3 cells were used as a positive control for HLA-G expression.

[0104] Figure 26: Mean Tumor Volume by Treatment Group. The mean (SEM) tumor volume of each treatment group is plotted (n=9). A 2-way ANOVA with a Dunnett’s multiple comparison test) was performed to compare treatment groups to Isotype Control. P values are presented as * color coordinated with the group that is being compared to Isotype control;*P < 0.05, **P < 0.01 , ***P < 0.001.

[0105] Figure 27: Mean change (A) in tumor volume in by treatment group. The mean ±SEM A tumor volume during the dosing period (Days 0-17) of each treatment group is plotted (n=9). A 2-way ANOVA with a Dunnett’s multiple comparison test) was performed to compare treatment groups to Isotype Control. P values are presented as * color coordinated with the group that is being compared to Isotype control; *P < 0.05, **P < 0.01 , ***P < 0.001.

[0106] Figure 28: CD8+ T cell Subsets in Tumor samples. Absolute count result (top) and percentage (bottom) of HLA-DR, CD69+, and PD-1+ cells expressed on CD8+ T cell subsets in tumor samples per flow cytometry analysis. P<0.05 is considered statistically significant compared with “Isotype group”.

[0107] DETAILED DESCRIPTION

[0108] Various preferred features and embodiments of the present invention will now be described by way of non-limiting examples.

[0109] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0110] The terms "comprising", "comprises" and "comprised of" as used herein are synonymous with "including", "includes", "containing", or "contains", and are inclusive or open-ended and do not exclude additional, non-recited members, elements or steps. The terms "comprising", "comprises" and "comprised of" also include the term "consisting of".

[0111] Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, any nucleic acid sequences are written left to right in 5' to 3' orientation and amino acid sequences are written left to right in amino to carboxy orientation, respectively.

[0112] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that such publications constitute prior art to the claims appended hereto. All publications mentioned in the specification are herein incorporated by reference.

[0113] This disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of this disclosure. The skilled person will understand that they can combine all features of the invention disclosed herein without departing from the scope of the invention as disclosed.

[0114] Sequence of antibody or antigen-binding fragment thereof

[0115] The present invention provides an antibody, or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises three complementarity determining regions on the heavy chain variable region (CDRHs), wherein the CDRHs have an amino acid sequence according to any one of SEQ ID NO: 1 to SEQ ID NO: 127, and wherein the antibody, or antigenbinding fragment thereof, comprises three complementarity determining regions on the light chain variable region (CDRLs), wherein the CDRLs have an amino acid sequence according to any one of SEQ ID NO: 128 to SEQ ID NO: 238. In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising:

[0116] (a) a CDRH1 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 37;

[0117] (b) a CDRH2 selected from the group consisting of SEQ ID NO 38 to SEQ ID NO 77; and

[0118] (c) a CDRH3 selected from the group consisting of SEQ ID NO 78 to SEQ ID NO 127; and a light chain variable region comprising:

[0119] (d) a CDRL1 selected from the group consisting of SEQ ID NO: 128 to SEQ ID NO: 168;

[0120] (e) a CDRL2 selected from the group consisting of SEQ ID NO: 169 to SEQ ID NO: 192; and

[0121] (f) a CDRL3 selected from the group consisting of SEQ ID NO: 193 to SEQ ID NO: 238.

[0122] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, wherein the heavy chain variable region comprises CDRs wherein CDRH1 , CDRH2 and CDRH3 are according to any one of the following groups of SEQ ID NOs in Table 1 :

[0123]

[0124] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, wherein the light chain variable region comprises CDRLs wherein CDRL1 , CDRL2 and CDRL3 are according to any one of the following groups of SEQ ID NOs in Table 2:

[0125] In one embodiment, the antibody, or antigen-binding fragment thereof, of the present invention may comprise CDRH1 , CDRH2 and CDRH3 as defined by the SEQ ID NOs of any of groups 1 - 50 of Table 1 and CDRL1 , CDRL2 and CDRL3 as defined by the SEQ ID NOs of any of group 1 - 50 of Table 2.

[0126] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, wherein the heavy chain variable region comprises or consists of amino acids according to any one of the following SEQ ID NOs in Table 3:

[0127] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, wherein the light chain variable region comprises or consists of amino acids according to any one of the following SEQ ID NOs in Table 4:

[0128] The KB identifiers provided in Tables 1-4 correspond to the KB identifiers provided in the Figures and Examples. For example, KB17A.5 sequences relate to KB17A.5.3 in the figures.

[0129] In one embodiment, the antibody, or antigen-binding fragment thereof, of the present invention comprises a heavy chain variable region as defined by any one of SEQ ID NOs: 239 to 288 and a light chain variable region as defined by any of SEQ ID NOs: 289 to 338.

[0130] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 1 , 38 and 78 and light chain variable region comprising CDRLs according to SEQ ID NO: 128, 169 and 193.

[0131] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 239, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 289. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 2,

[0132] 39 and 79 and a light chain variable region comprising CDRLs according to SEQ ID NO: 129, 170 and 194.

[0133] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 240, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 290.

[0134] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 3,

[0135] 40 and 80 and a light chain variable region comprising CDRLs according to SEQ ID NO: 130, 171 and 195.

[0136] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 241 , and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 291.

[0137] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 9, 74 and 123 and a light chain variable region comprising CDRLs according to SEQ ID NO: 143, 190 and 234.

[0138] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 285, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 335.

[0139] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 6, 43 and 83 and a light chain variable region comprising CDRLs according to SEQ ID NO: 133, 178 and 198.

[0140] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 247, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 297.

[0141] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 16, 52 and 93 and a light chain variable region comprising CDRLs according to SEQ ID NO: 142, 171 and 207. 1 In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 256, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 306.

[0142] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 19, 54 and 96 and a light chain variable region comprising CDRLs according to SEQ ID NO: 145, 182 and 210.

[0143] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 259, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 309.

[0144] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 8, 44 and 85 and a light chain variable region comprising CDRLs according to SEQ ID NO: 135, 175 and 200.

[0145] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO 249, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 299.

[0146] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 12, 48 and 89 and a light chain variable region comprising CDRLs according to SEQ ID NO: 139, 179 and 203.

[0147] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 252, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 302.

[0148] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 64 and 107 and a light chain variable region comprising CDRLs according to SEQ ID NO: 154, 169 and 220.

[0149] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 269, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 319.

[0150] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 33, 54 and 118 and a light chain variable region comprising CDRLs according to SEQ ID NO: 143, 181 and 229.

[0151] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 280, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 330.

[0152] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 40 and 121 and a light chain variable region comprising CDRLs according to SEQ ID NO: 165, 171 and 232.

[0153] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 283, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 333.

[0154] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 76 and 125 and a light chain variable region comprising CDRLs according to SEQ ID NO: 143, 171 and 236.

[0155] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 286, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 336.

[0156] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 37, 40 and 126 and a light chain variable region comprising CDRLs according to SEQ ID NO: 162, 191 and 237.

[0157] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 287, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 337. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 11 , 47 and 88 and a light chain variable region comprising CDRLs according to SEQ ID NO: 138, 178 and 202.

[0158] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 242, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 292.

[0159] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 36, 75 and 124 and a light chain variable region comprising CDRLs according to SEQ ID NO: 167, 174 and 235.

[0160] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 243, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 293.

[0161] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 22, 57 and 100 and a light chain variable region comprising CDRLs according to SEQ ID NO: 148, 180 and 214.

[0162] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 244, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 294.

[0163] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 4,

[0164] 41 and 81 and a light chain variable region comprising CDRLs according to SEQ ID NO: 131 , 172 and 196.

[0165] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 245, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 295.

[0166] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 5,

[0167] 42 and 82 and a light chain variable region comprising CDRLs according to SEQ ID NO: 132, 173 and 197. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 246, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 296.

[0168] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 7,

[0169] 44 and 84 and a light chain variable region comprising CDRLs according to SEQ ID NO: 134, 170 and 199.

[0170] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 248, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 298.

[0171] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 9,

[0172] 45 and 86 and a light chain variable region comprising CDRLs according to SEQ ID NO: 136, 176 and 196.

[0173] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 250, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 300.

[0174] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 10, 46 and 87 and a light chain variable region comprising CDRLs according to SEQ ID NO: 137, 177 and 201.

[0175] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 251 , and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 301.

[0176] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 49 and 90 and a light chain variable region comprising CDRLs according to SEQ ID NO: 140, 173 and 204.

[0177] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 253, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 303.

[0178] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 14,

[0179] 50 and 91 and a light chain variable region comprising CDRLs according to SEQ ID NO: 138, 178 and 205.

[0180] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 254, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 304.

[0181] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 15,

[0182] 51 and 92 and a light chain variable region comprising CDRLs according to SEQ ID NO: 141 , 171 and 206.

[0183] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 255, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 305.

[0184] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 17, 44 and 94 and a light chain variable region comprising CDRLs according to SEQ ID NO: 143, 181 and 208.

[0185] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 257, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 307.

[0186] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 18, 53 and 95 and a light chain variable region comprising CDRLs according to SEQ ID NO: 144, 171 and 209.

[0187] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 258, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 308. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 20,

[0188] 55 and 97 and a light chain variable region comprising CDRLs according to SEQ ID NO: 138, 169 and 211.

[0189] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 260, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 310.

[0190] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 21 ,

[0191] 56 and 98 and a light chain variable region comprising CDRLs according to SEQ ID NO: 146, 170 and 212.

[0192] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 261 , and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 311.

[0193] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 40 and 99 and a light chain variable region comprising CDRLs according to SEQ ID NO: 147, 169 and 213.

[0194] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 262, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 312.

[0195] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 23,

[0196] 58 and 101 and a light chain variable region comprising CDRLs according to SEQ ID NO: 143, 181 and 215.

[0197] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 263, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 313.

[0198] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 24,

[0199] 59 and 102 and a light chain variable region comprising CDRLs according to SEQ ID NO: 149, 183 and 201. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 264, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 314.

[0200] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 25,

[0201] 60 and 103 and a light chain variable region comprising CDRLs according to SEQ ID NO: 150, 169 and 216.

[0202] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 265, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 315.

[0203] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 26,

[0204] 61 and 104 and a light chain variable region comprising CDRLs according to SEQ ID NO: 151 , 184 and 217.

[0205] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 266, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 316.

[0206] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 27,

[0207] 62 and 105 and a light chain variable region comprising CDRLs according to SEQ ID NO: 152, 169 and 218.

[0208] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 267, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 317.

[0209] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 28,

[0210] 63 and 106 and a light chain variable region comprising CDRLs according to SEQ ID NO: 153, 185 and 219.

[0211] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 268, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 318.

[0212] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 29,

[0213] 65 and 108 and a light chain variable region comprising CDRLs according to SEQ ID NO: 155, 171 and 221.

[0214] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 270, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 320.

[0215] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 9,

[0216] 66 and 109 and a light chain variable region comprising CDRLs according to SEQ ID NO: 138, 169 and 201.

[0217] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 271 , and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 321.

[0218] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 23,

[0219] 67 and 110 and a light chain variable region comprising CDRLs according to SEQ ID NO: 156, 186 and 222.

[0220] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 272, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 322.

[0221] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 30,

[0222] 68 and 111 and a light chain variable region comprising CDRLs according to SEQ ID NO: 157, 171 and 223.

[0223] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 273, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 323. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 69 and 112 and a light chain variable region comprising CDRLs according to SEQ ID NO: 158, 171 and 224.

[0224] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 274, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 324.

[0225] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 9, 44 and 113 and a light chain variable region comprising CDRLs according to SEQ ID NO: 159, 171 and 225.

[0226] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 275, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 325.

[0227] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 31 , 70 and 114 and a light chain variable region comprising CDRLs according to SEQ ID NO: 160, 187 and 226.

[0228] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 276, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 326.

[0229] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 32, 66 and 115 and a light chain variable region comprising CDRLs according to SEQ ID NO: 161 , 188 and 227.

[0230] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 277, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 327.

[0231] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 23, 44 and 116 and a light chain variable region comprising CDRLs according to SEQ ID NO: 162, 171 and 209. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 278, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 328.

[0232] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 9, 63 and 117 and a light chain variable region comprising CDRLs according to SEQ ID NO: 163, 170 and 228.

[0233] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 279, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 329.

[0234] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 34, 71 and 119 and a light chain variable region comprising CDRLs according to SEQ ID NO: 162, 186 and 230. In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 281 , and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 331.

[0235] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 6,

[0236] 72 and 120 and a light chain variable region comprising CDRLs according to SEQ ID NO: 164, 189 and 231.

[0237] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 282, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 332.

[0238] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 35,

[0239] 73 and 122 and a light chain variable region comprising CDRLs according to SEQ ID NO: 166, 173 and 233.

[0240] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 284, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 334.

[0241] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising CDRHs according to SEQ ID NO: 13, 77 and 127 and a light chain variable region comprising CDRLs according to SEQ ID NO: 168, 192 and 238.

[0242] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 288, and a light chain variable region comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 338.

[0243] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to the following sequence:

[0244] QVQLVQSGAEVKKPGASVKVSCKASGYTFISNYMHWVRQAPGQGLEWMGLINPSGGSTIYAQK

[0245] FQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCLTGRSWGQGTLVTVSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCW VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 339)

[0246] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a light chain comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to the following sequence:

[0247] DIQMTQSPSSLSASVGDRVTITCRASQGISSYLAWYQQKPGKAPKLLIYKASSLESGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCLQSYSVPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC (SEQ ID NO: 340)

[0248] In one embodiment, the antibody, or antigen-binding fragment thereof, according to the present invention comprises a heavy chain comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 339, and a light chain comprising an amino acid sequence having at least about 70% identity, such as at least about 75% identity, such as at least about 80% identity, such as at least about 85% identity, such as at least about 90% identity, such as at least about 95% identity, such as at least about 96% identity, such as at least about 97% identity, such as at least about 98% identity, such as at least about 99% identity, such as 100% identity to SEQ ID NO: 340.

[0249] Antibody Binding Properties

[0250] The antibody of the invention, or antigen-binding fragment thereof, may be defined by its binding specificity. In one embodiment, the antibody, or antigen-binding fragment thereof, binds specifically to HLA-G. In other words, the antibody, or antigen-binding fragment thereof, does not bind to HLA-A, HLA-B, HLA-C, HLA-E or HLA-F.

[0251] Binding specificity may be measured through a HLA-G On-Target Screen and a HLA-G Off-Target Screen as detailed in the Examples.

[0252] The antibody of the invention, or antigen-binding fragment thereof, may be defined by its EC50 value. The EC50 value is a measure of the half-maximal effective concentration, indicating the concentration of a drug required to have half the maximum effect. The binding of the antibodies, or antigen-binding fragments thereof, disclosed herein may therefore be represented through their EC50 value as determined by any suitable assay, such as an ELISA.

[0253] Direct ELISA is used herein to measure the EC50 of anti-HLA-G mAbs to recombinant human HLAG complex. Suitably, an antibody, or antigen-binding fragment thereof, disclosed herein, may have an EC50 value, as measured by direct ELISA, of about 20 pg / mL or less, about 15 pg / mL or less, about 10 pg / mL or less, about 5 pg / mL or less, about 2 pg / mL or less, about 1 pg / mL or less, about 0.5 pg / mL or less, about 0.4 pg / mL or less, about 0.3 pg / mL or less, about 0.2 pg / mL or less, about 0.15 pg / mL or less, about 0.1 pg / mL or less, or about 0.05 pg / mL or less.

[0254] Suitably, an antibody, or antigen-binding fragment thereof, disclosed herein, may have an EC50 value, as measured by direct ELISA, of about 0.2 pg / mL or less.

[0255] Suitably, an antibody, or antigen-binding fragment thereof, disclosed herein, may have an EC50 value, as measured by direct ELISA, of about 0.15 pg / mL or less.

[0256] Suitably, an antibody, or antigen-binding fragment thereof, disclosed herein, may have an EC50 value, as measured by direct ELISA, of about 0.1 pg / mL or less.

[0257] Suitably, an antibody, or antigen-binding fragment thereof, disclosed herein, may have an EC50 value, as measured by direct ELISA, of about 0.05 pg / mL or less.

[0258] Antibody binding may also be measured through a competitive ELISA. This assay is used herein to measure how much the anti-HLA-G antibodies disclosed herein may block the binding of recombinant HLA-G to recombinant LILRB2. Blocking may be interpreted herein as preventing or inhibiting HLA-G and LILRB2 from binding and / or forming a complex. The capability of the antibodies disclosed herein to block such an interaction may be represented as a % blocking. The higher the percent blocking the better the capacity of the mAb to block binding of the two recombinant antigens.

[0259] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70% or at least about 80% blocking of HLA-G and LILRB2 interaction.

[0260] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70% or at least about 80% blocking of HLA-G and LILRB2 interaction, at a given concentration of HLA-G or LILRB2.

[0261] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 20% blocking of HLA-G and LILRB2 interaction.

[0262] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 30% blocking of HLA-G and LILRB2 interaction.

[0263] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 40% blocking of HLA-G and LILRB2 interaction.

[0264] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 50% blocking of HLA-G and LILRB2 interaction.

[0265] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 60% blocking of HLA-G and LILRB2 interaction.

[0266] Antibody Fc Properties

[0267] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a neonatal fragment crystallizable Fc receptor (FcRn).

[0268] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a human FcRn receptor. In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to an Fc receptor.

[0269] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a human Fc receptor.

[0270] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a human CD64.

[0271] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a human CD16a.

[0272] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a human CD32b / c.

[0273] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein, can bind to a human CD32a.

[0274] Suitably, in some embodiments, antibody, or antigen-binding fragment thereof, binding to an Fc receptor, such a human Fc receptor, promotes macrophage and NK cell mediated antibodydependent cellular cytotoxicity (ADCC) activity.

[0275] Antibody Thermostability

[0276] The antibody, or antigen-binding fragment thereof, may be further defined by its thermostability. Thermostability may be defined by the unfolding events that occur as the antibody, or antigenbinding fragment thereof, experiences increasing temperatures. ‘Tonset’ is the temperature at which unfolding events begin to occur, ‘Tm’ is the temperature at the midpoint of the unfolding events, and Tagg is the temperature at which the unfolding events have resulted in aggregation. ‘Tm’, the midpoint of unfolding events, usually occurs as two transitions referred to as Tm1 and Tm2. The Tm1 value represents the first transition which is thought to involve the thermal unfolding of the CH2 domain. The Tm2 value represents a second transition which is representative of the unfolding of the CH3 / Fab domain.

[0277] In one embodiment, the antibody, or antigen-binding fragment thereof, is thermostable up 60°C.

[0278] In one embodiment, the antibody, or antigen-binding fragment thereof, is thermostable up 70°C. In one embodiment, the antibody, or antigen-binding fragment thereof, is thermostable up 80°C.

[0279] Additional features of antibody or antigen-binding fragment thereof

[0280] In one embodiment, the antibody, or antigen-binding fragment thereof, may be IgG and all isotypes thereof.

[0281] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG1 and all allotypes thereof.

[0282] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG2 and all allotypes thereof.

[0283] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG3 and all allotypes thereof.

[0284] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG4 and all allotypes thereof.

[0285] In one embodiment the antibody, or antigen-binding fragment thereof, may be IgA and all isotypes thereof.

[0286] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgA1 and allotypes thereof.

[0287] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgA2 and all allotypes thereof.

[0288] In one embodiment the antibody, or antigen-binding fragment thereof, may be IgM and all allotypes thereof.

[0289] In one embodiment the antibody, or antigen-binding fragment thereof, may be IgE and all allotypes thereof.

[0290] In one embodiment the antibody, or antigen-binding fragment thereof, may be IgD and all isotypes thereof. In one embodiment the antibody, or antigen-binding fragment thereof, may be lgD1 and all allotypes thereof.

[0291] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgD2 and all allotypes thereof.

[0292] In one embodiment, the antibody, or antigen-binding fragment thereof is a chimeric antibody.

[0293] In one embodiment, the antibody, or antigen-binding fragment thereof, is a humanized antibody.

[0294] In one embodiment, the antibody, or antigen-binding fragment thereof is an affinity-matured antibody.

[0295] In one embodiment, the antibody, or antigen-binding fragment thereof, is an affinity-matured, humanized antibody.

[0296] Accordingly, the terms “humanized”, “humanization”, or other forms related to the term “humanize” when used in the context of this disclosure shall be understood as referring to an antibody structure, which may be a complete antibody or any portion or fragment thereof, which are modified to closely resemble the human lgG1 but retaining at least one CDR of non-human origin. Such modifications may be performed on a murine IgG antibody, for example lgG1 , and in some embodiments, the subject monoclonal antibodies are humanized or chimeric. Such modifications can be characterized by a substitution of one or more amino acids consistent with a human germline sequence for a corresponding number of amino acids found in a non-human mammalian form of the lgG1. Such substitutions may occur in one or more of the CDRs or within the framework regions between the CDRs, or indeed anywhere on the light and heavy variable chains.

[0297] Antibody fragments

[0298] The scope of embodiments disclosed herein includes all naturally occurring antibodies, such as IgG, IgM, IgA, IgE and IgD and their respective subtypes and allotypes. The scope of embodiments further includes antigen-binding fragments of said antibody, including but not limited to an antigenbinding fragment (Fab), a fragment antibody (F(ab’)2), single chain antibody (scFv) and a singledomain antibody (sdAb). A fragment antibody (F(ab’)2), refers to a region on an antibody that remains following digestion of the Fc region which leaving intact portions of the hinge region.

[0299] A single chain antibody (scFv) refers to an antibody that has been engineered to consist of a light and heavy chain variable region that are connected by a peptide-linker sequence. The peptide linker sequence is typically in the length of 10-25 amino acids, rich in glycine for flexibility and rich in serine or threonine for solubility. The peptide linker may connect the N-terminus of the heavy chain variable region with the C-terminus of the light chain variable region.

[0300] A single domain antibody (sdAb), which is often referred to as a nanobody, refers to an antigenbinding fragment of an antibody that consists of a single monomeric variable antibody domain. Therefore, an sdAb may be a light chain variable region or a heavy chain variable region. Examples of single-domain antibodies include, but are not limited to, VHH fragments and VNAR fragments. VHH and VNAR fragments comprise the antigen-binding fragment of the heavy chain.

[0301] In one embodiment, the antigen-binding fragment of the antibodies disclosed herein may be any fragment of the antibodies disclosed herein.

[0302] In another embodiment the antigen-binding fragment of the antibodies disclosed herein may be a genetically-engineered product of one or more of the fragments of the antibody.

[0303] Whilst the disclosure herein discusses IgG at greater length, the scope of embodiments is not limited to IgG, but includes other naturally occurring antibodies, such as IgM, IgA, IgE and IgD and their respective subtypes.

[0304] In one embodiment the fragment is an antigen-binding fragment (Fab), a fragment antibody (F(ab’)2), a single-chain variable fragment (scFV) or a single domain antibody (sdAb), or a camelid antibody (VHH).

[0305] Pharmaceutical formulations

[0306] The antibody, or antigen binding fragment thereof, disclosed herein may be interpreted throughout this description as an antibody, or antigen-binding fragment thereof, or a pharmaceutically acceptable salt thereof. The antibody, or antigen binding fragment thereof, disclosed herein may be interpreted throughout this description as a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein.

[0307] In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, of the present invention. The antibody, or antigenbinding fragment thereof may be in combination with a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0308] Medical use

[0309] The present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, or pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein, for use in therapy.

[0310] The present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, or pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein, for use as a medicament.

[0311] The present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein, or pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein, for the manufacture of a medicament.

[0312] The present invention provides a method of treatment comprising administering an antibody, or antigen-binding fragment thereof, as disclosed herein, or pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein, to a subject in need thereof.

[0313] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein for use as an anti-cancer therapeutic.

[0314] In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein, for use as an anti-cancer therapeutic.

[0315] In one embodiment, the present invention provides the antibody, or antigen-binding fragment thereof, as disclosed herein for use in the treatment or prevention of cancer. In one embodiment, the present invention provides the pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein for use in the treatment or prevention of cancer.

[0316] In one embodiment, the present invention provides a method of treatment of a cancer comprising administering an antibody, or antigen-binding fragment thereof, as disclosed herein.

[0317] In one embodiment, the present invention provides a method of treatment of a cancer comprising administering a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein.

[0318] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein for use in the production of an immunogenic response.

[0319] In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein for use in the production of an immunogenic response.

[0320] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein for use in preventing the induction of immunosuppressive pathways.

[0321] In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein for use in preventing the induction of immunosuppressive pathways.

[0322] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein for use in preventing the induction of immunosuppressive pathways due to the interaction between HLA-G and LILRB2.

[0323] In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein for use in preventing the induction of immunosuppressive pathways due to the interaction between HLA-G and LILRB2.

[0324] In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein for use in preventing the induction of immunosuppressive pathways due to the interaction between HLA-G expressed on tumour and pathogenic cells and LILRB2 expressed on immune cells. In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, as disclosed herein for use in preventing the induction of immunosuppressive pathways due to the interaction between HLA-G expressed on tumour and pathogenic cells and LILRB2 expressed on immune cells.

[0325] LILRB2 may be expressed on immune cells, such as myeloid cells.

[0326] LILRB2 may be expressed on immune cells, such as monocytes, macrophages, granulocytes.

[0327] LILRB2 may be expressed on immune cells such as, T-cells.

[0328] In some embodiments, the subject of the medical uses and methods of treatment according to the present invention may be a mammal.

[0329] In some embodiments, the subject may be a human.

[0330] In some embodiments, the subject may be a patient, such as a human patient.

[0331] In some embodiments, the subject may suffer from and / or have been diagnosed with cancer.

[0332] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the prevention of HLA-G induced monocyte suppression.

[0333] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of cancer.

[0334] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of solid tumours.

[0335] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of bladder cancer.

[0336] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of placental cancer. In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of melanoma.

[0337] Antibody-drug conjugates

[0338] Antibody-drug conjugates (ADCs) are therapeutic molecules that may be specifically targeted to an antigen via the antibody, or an antigen-binding fragment thereof, enabling precise delivery of a drug to a specific location. For example, ADCs have been used as chemotherapeutic agents to target cytotoxic drugs to antigen-expressing tumour cells, whereupon they are internalized by said tumour cells, enabling the cytotoxic drug to take effect in specific tumour cells. This strategy has been further advanced through the conjugation of drugs to antibodies through cleavable linker molecules, enabling the ADC to be cleaved once internalised into a specific cell, releasing the cytotoxic drug such that it has greater activity.

[0339] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein.

[0340] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety.

[0341] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety connected by a cleavable linker-molecule.

[0342] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety connected by a non-cleavable linker-molecule.

[0343] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety wherein the coupling moiety is a drug.

[0344] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety wherein the coupling moiety is a toxin. In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety wherein the coupling moiety is a cytokine.

[0345] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety wherein the coupling moiety is an enzyme.

[0346] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein and a coupling moiety wherein the coupling moiety is a drug, toxin, cytokine, enzyme or a combination thereof.

[0347] Antibody-Toxin Fusion Proteins

[0348] An anti body- toxin fusion protein, such as a recombinant immunotoxin, comprises an antibody, or antigen-binding fragment thereof, or immunoglobulin that is linked to a peptide toxin. Such an anti body- toxin fusion protein may be encoded by a recombinant DNA sequence that is subsequently expressed. The recombinant DNA sequence, also known as a fusion gene, may be expressed via an expression vector or plasmid that comprises the DNA segments which thus direct the synthesis of such a fusion protein. The anti body- toxin fusion protein, or recombinant immunotoxin, therefore comprises an amino acid sequence representing an antibody, antigen-binding fragment thereof, or immunoglobulin linked to peptide toxin.

[0349] In other words, a recombinant immunotoxin is a polypeptide in which a peptide toxin is genetically linked to an antibody component, often by a contiguous polypeptide linker.

[0350] Much like antibody-drug conjugates, anti body- toxin fusion proteins enable targeted accumulation and activation of the toxin for specific effect, promoting efficiency while minimising harm to normal cells.

[0351] In one embodiment, the present invention provides a recombinant fusion protein wherein the fusion protein comprises an antibody, or antigen-binding fragment thereof, disclosed herein that is linked to a peptide toxin.

[0352] In one embodiment, the present invention provides a recombinant fusion protein wherein the fusion protein comprises an antibody, or antigen-binding fragment thereof, as disclosed herein, wherein at least one of the heavy or light chain regions are linked to a peptide toxin. Nucleotide Sequences

[0353] The antibody, or antigen-binding fragment thereof, may be encoded by any suitable nucleotide sequence.

[0354] A person skilled in the art will immediately understand that any antibody, or antigen binding fragment as disclosed herein, may be encoded by an appropriate nucleotide sequence.

[0355] In some embodiments, the nucleotide sequence is codon-optimised, for example codon-optimised for expression in humans or plants. Cells may differ in their bias towards particular codons, some of which are used more. This codon bias corresponds to the relative abundance of particular tRNAs in a given cell type. As such, codons may be altered in a sequence such that they match with the tRNA with the greatest relative abundance in a cell type, thereby increasing expression. Similarly, by altering the sequence such that it matches the least abundant tRNA, one can reduce expression. Therefore, a greater degree of translational control is available. Codon usage tables are known in the art for a variety of organisms.

[0356] In one embodiment, the present invention provides a nucleic acid that encodes an antibody, or antigen-binding fragment thereof, as disclosed herein.

[0357] In one embodiment, the present invention provides a nucleic acid that encodes the heavy chain variable region and light chain variable region an antibody, or antigen-binding fragment thereof, as disclosed herein.

[0358] In one embodiment, the present invention provides a nucleic acid that encodes heavy chain variable region as defined by the amino acid sequence of any one of SEQ ID Nos: 239 to 288 and a light chain variable region as defined by the amino acid sequence of any of SEQ ID Nos: 289 to 338.

[0359] Expression System

[0360] The present invention further relates to a method of production and engineering of antibodies, or antigen-binding fragments thereof, disclosed herein.

[0361] In some embodiments, the present invention provides a method of expressing a nucleic acid encoding the antibody, or antigen binding fragment thereof, as disclosed herein, in a cell or cell-free expression system. In some embodiments, a range of production hosts may be used for production of the antibody, or an antigen-binding fragment thereof, disclosed herein. Such hosts, which are utilized for the production of antibodies, or antigen-binding fragments thereof, may include, but are not limited to mammalian cells (eg. CHO cells), plant cells (eg. Nicotiana benthamiana cells), eukaryotic cells (eg. Yeast cells) and microbial cells, such as bacterial cells. Suitable bacterial hosts include, but are not limited to, Escherichia (e.g. E. coli), Bacillus (e.g. Bacillus subtilis), Mycobacterium (e.g. M. bovis BCG) and Salmonella.

[0362] In some embodiments within the scope of the present disclosure and as known in the art, one or more plasmid expression vectors may be assembled and transfected into living cells for transient expression by the host. If multiple expression vectors are used, they can be transfected into the cells either separately or jointly, and expressed by the host. In a preferred embodiment, an engineered Nicotiana benthamiana (Nb) plant strain is the production host into which one or more expression vectors containing nucleic acids encoding the antibody, or antigen-binding fragments thereof, of the present disclosure are transfected.

[0363] In one embodiment, the plasmid expression vector is transfected into Agrobacterium hosts. In a preferred embodiment, the plasmid expression vector is transfected into Agrobacterium tumefaciens.

[0364] In a further embodiment, vector-carrying Agrobacterium are introduced into plant hosts by spray inoculation in the presence of mild abrasion. In a further embodiment, vector-carrying Agrobacterium are introduced into plant hosts by vacuum infiltration.

[0365] Further Embodiments

[0366] Monoclonal antibodies, including immunoglobulins, have been engineered and produced in various kinds of eukaryotic cells and other living cells. However, mammalian expression systems tend to be hampered by slow production turnaround cycles, and production through the use of certain cell lines may be constrained by the presence of mammalian viruses and other undesired compounds produced alongside the monoclonal antibodies in mammalian systems.

[0367] While not intending to limit the scope of the present embodiments, it is noted that plant based antibody production systems can avoid certain pitfalls of mammalian-based monoclonal antibody production. One such system, provided as a non-limiting example, utilizes an engineered Nicotiana benthamiana (Nb) plant strain expressing one or more expression vectors containing nucleic acid constructs for the formation of the desired mAbs. In some embodiments within the scope of the present disclosure, and as known in the art, one or more plasmid expression vectors are assembled and then transfected into living cells for transient expression by the host. If multiple expression vectors are used, they can be transfected into the cells, for example as found in plant tissue of Nb, either separately or jointly, and expressed by the host. An exemplary transient expression process uses Agrobacterium strains (e.g., Agrobacterium tumefaciens), in which the Agrobacterium are grown and infiltrated into whole plants, infecting the plant and introducing the genes to the plant cells. After infiltration and transient expression of the genes of interest in the germinated plants, proper feeding and care of the infiltrated plants provides for desired plant growth and ultimately harvesting of the mAb-containing infiltrated plant samples. The desired mAb product then is extracted from the harvested parts of the plant, generally followed by purification and characterization of the extracted monoclonal antibodies.

[0368] While the manner of introducing the vector-carrying Agrobacterium into these hosts is not intended as a limiting factor, spray inoculation in the presence of mild abrasion and vacuum infiltration are options. Again, other methods can be employed with plant hosts, and other non-plant hosts can be utilized with the nucleic acid constructs introduced to those hosts through known methods.

[0369] As will be appreciated by persons of ordinary skill in the relevant art, a range of other production hosts can be suitable for use in mAb production. Such hosts which are utilized for the production of mAbs in accordance with the present embodiments may include, but are not necessarily limited to, mammalian (e.g., CHO cells), plant (E.g., Nb plants or plant cells), yeast, and eukaryotic, and microbial cells. Through known steps available in the relevant literature, such hosts have been developed through genetic engineering, mutagenesis, or (in the case of plants) selective breeding aimed at enhancing mAb production or provide additional physicochemical characteristics in terms of increased or decreased molecular, metabolic, chemical, phenotypic, or other traits that affect protein formation by host.

[0370] Numbered paragraphs

[0371] 1. An antibody, or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising:

[0372] (a) a CDRH1 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 37;

[0373] (b) a CDRH2 selected from the group consisting of SEQ ID NO 38 to SEQ ID NO 77; and

[0374] (c) a CDRH3 selected from the group consisting of SEQ ID NO 78 to SEQ ID NO 127; and a light chain variable region comprising: (d) a CDRL1 selected from the group consisting of SEQ ID NO: 128 to SEQ ID NO: 168;

[0375] (e) a CDRL2 selected from the group consisting of SEQ ID NO: 169 to SEQ ID NO: 192; and

[0376] (f) a CDRL3 selected from the group consisting of SEQ ID NO: 193 to SEQ ID NO: 238.

[0377] 2. The antibody, or antigen-binding fragment thereof, of paragraph 1, wherein the heavy chain variable region comprise CDRs selected from one of the following:

[0378] (a) CDRHs according to SEQ ID NO: 1, 38 and 78;

[0379] (b) CDRHs according to SEQ ID NO: 2, 39 and 79;

[0380] (c) CDRHs according to SEQ ID NO: 3, 40 and 80;

[0381] (d) CDRHs according to SEQ ID NO: 9, 74 and 123;

[0382] (e) CDRHs according to SEQ ID NO: 6, 43 and 83;

[0383] (f) CDRHs according to SEQ ID NO: 16, 52 and 93;

[0384] (g) CDRHs according to SEQ ID NO: 19, 54 and 96;

[0385] (h) CDRHs according to SEQ ID NO: 8, 44 and 85;

[0386] (i) CDRHs according to SEQ ID NO: 12, 48 and 89;

[0387] (j) CDRHs according to SEQ ID NO: 13, 64 and 107;

[0388] (k) CDRHs according to SEQ ID NO: 33, 54 and 118;

[0389] (l) CDRHs according to SEQ ID NO: 13, 40 and 121 ;

[0390] (m) CDRHs according to SEQ ID NO: 13, 76 and 125; or

[0391] (n) CDRHs according to SEQ ID NO: 37, 40 and 126.

[0392] 3. The antibody, or antigen-binding fragment thereof, of paragraph 1 or 2, wherein the light chain variable region comprises CDRs selected from one of the following:

[0393] (a) CDRLs according to SEQ ID NO: 128, 169 and 193;

[0394] (b) CDRLs according to SEQ ID NO: 129, 170 and 194;

[0395] (c) CDRLs according to SEQ ID NO: 130, 171 and 195

[0396] (d) CDRLs according to SEQ ID NO: 143, 190 and 234;

[0397] (e) CDRLs according to SEQ ID NO: 133, 178 and 198;

[0398] (f) CDRLs according to SEQ ID NO: 142, 171 and 207;

[0399] (g) CDRLs according to SEQ ID NO: 145, 182 and 210;

[0400] (h) CDRLs according to SEQ ID NO: 135, 175 and 200;

[0401] (i) CDRLs according to SEQ ID NO: 139, 179 and 203;

[0402] (j) CDRLs according to SEQ ID NO: 154, 169 and 220;

[0403] (k) CDRLs according to SEQ ID NO: 143, 181 and 229; (l) CDRLs according to SEQ ID NO: 165, 171 and 232;

[0404] (m) CDRLs according to SEQ ID NO: 143, 171 and 236; or

[0405] (n) CDRLs according to SEQ ID NO: 162, 191 and 237.

[0406] 4. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 1 , 38 and 78 and CDRLs according to SEQ ID NO: 128, 169 and 193.

[0407] 5. The antibody, or antigen-binding fragment thereof, according to paragraph 4, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 239, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 289.

[0408] 6. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 2, 39 and 79 and CDRLs according to SEQ ID NO: 129, 170 and 194.

[0409] 7. The antibody, or antigen-binding fragment thereof, according to paragraph 6, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 240, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 290.

[0410] 8. The antibody, or antigen-binding fragment thereof, according to paragraph 6 or paragraph 7, wherein the heavy chain comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 339 and wherein the light chain comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 340.

[0411] 9. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 3, 40 and 80 and CDRLs according to SEQ ID NO: 130, 171 and 195.

[0412] 10. The antibody, or antigen-binding fragment thereof, according to paragraph 9, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 241 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 291. 11. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 9, 74 and 123 and CDRLs according to SEQ ID NO: 143, 190 and 234.

[0413] 12. The antibody, or antigen-binding fragment thereof, according to paragraph 11 , wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 285, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 335.

[0414] 13. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 6, 43 and 83 and CDRLs according to SEQ ID NO: 133, 178 and 198.

[0415] 14. The antibody, or antigen-binding fragment thereof, according to paragraph 13, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 247, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 297.

[0416] 15. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 16, 52 and 93 and CDRLs according to SEQ ID NO: 142, 171 and 207.

[0417] 16. The antibody, or antigen-binding fragment thereof, according to paragraph 15, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 256, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 306.

[0418] 17. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 19, 54 and 96 and CDRLs according to SEQ ID NO: 145, 182 and 210.

[0419] 18. The antibody, or antigen-binding fragment thereof, according to paragraph 17, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 259, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 309. 19. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 8, 44 and 85 and CDRLs according to SEQ ID NO: 135, 175 and 200.

[0420] 20. The antibody, or antigen-binding fragment thereof, according to paragraph 19, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 249, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 299.

[0421] 21. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 12, 48 and 89 and CDRLs according to SEQ ID NO: 139, 179 and 203.

[0422] 22. The antibody, or antigen-binding fragment thereof, according to paragraph 21 , wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 252, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 302.

[0423] 23. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 64 and 107 and CDRLs according to SEQ ID NO: 154, 169 and 220.

[0424] 24. The antibody, or antigen-binding fragment thereof, according to paragraph 23, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 269, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 319.

[0425] 25. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 33, 54 and 118 and CDRLs according to SEQ ID NO: 143, 181 and 229.

[0426] 26. The antibody, or antigen-binding fragment thereof, according to paragraphs 25, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 280, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 330. 27. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 40 and 121 and CDRLs according to SEQ ID NO: 165, 171 and 232.

[0427] 28. The antibody, or antigen-binding fragment thereof, according to paragraph 27, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 283, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 333.

[0428] 29. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 76 and 125 and CDRLs according to SEQ ID NO: 143, 171 and 236.

[0429] 30. The antibody, or antigen-binding fragment thereof, according to paragraph 29, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO

[0430] 286, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 336.

[0431] 31. The antibody, or antigen-binding fragment thereof, according to any of paragraphs 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 37, 40 and 126 and CDRLs according to SEQ ID NO: 162, 191 and 237.

[0432] 32. The antibody, or antigen-binding fragment thereof, according to paragraph 31 , wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO

[0433] 287, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 337.

[0434] 33. The antibody, or antigen-binding fragment thereof, of any one of paragraphs 5, 7, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 or 32 wherein the antibody, or antigen-binding fragment thereof, has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% sequence identity to the recited sequence.

[0435] 34. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, wherein the antibody or antigen binding fragment is a single-chain variable fragment (scFV), a single-domain antibody (sdAb), an antigen-binding fragment (Fab), or a fragment antibody (F(ab’)2). 35. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, wherein the antibody or antigen-binding fragment is IgG, IgA, IgM, IgE or IgD isotype or any allotype thereof.

[0436] 36. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, wherein the antibody binds to HLA-G.

[0437] 37. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs for use in therapy.

[0438] 38. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, for use in the treatment or prevention of cancer.

[0439] 39. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, for use in the manufacture of a medicament for the treatment of cancer.

[0440] 40. A nucleic acid encoding an antibody, or an antigen-binding fragment thereof, according to any one of the preceding paragraphs.

[0441] 41. A pharmaceutical composition comprising the antibody, or antigen-binding fragment thereof, or nucleic acid of any one of the preceding paragraphs.

[0442] 42. A method of treating cancer comprising administering the antibody, or antigen-binding fragment thereof, the pharmaceutical composition or nucleic acid of any one of the preceding paragraphs.

[0443] 43. An antibody, or antigen-binding fragment thereof, pharmaceutical composition or nucleic acid of any one of the previous paragraphs for use in the production of an immunogenic response.

[0444] 44. An antibody, or antigen-binding fragment thereof, pharmaceutical composition or nucleic acid of any one of the previous paragraphs for use in preventing the induction of immunosuppressive pathways.

[0445] 45. An antibody, or antigen-binding fragment thereof, pharmaceutical composition or nucleic acid of any one of the previous paragraphs for use in preventing the induction of immunosuppressive pathways due to the interaction between HLA-G and LILRB2. 46. An antibody, or antigen-binding fragment thereof, pharmaceutical composition or nucleic acid of any one of the previous paragraphs for use in preventing the induction of immunosuppressive pathways due to the interaction between HLA-G expressed on tumour cells and LILRB2 expressed on immune cells, wherein the immune cell may be a myeloid cell.

[0446] EXAMPLES

[0447] 7. HLA-G On-Target Screen

[0448] HEK293T cells were transfected with plasmid encoding HLA-G using the Lipofectamine 3000. Two days after transfection, cells were harvested and stained sequentially with viability dye, primary antibody (mAb clone or human lgG1 isotype control) at 0.15 ug / ml, and then finally with anti-human IgG-APC at 5 ug / ml. A commercial anti-HLA-G-APC antibody was used as a positive control to demonstrate HLA-G expression on the HEK293T cell line after transfection. Cell suspensions were acquired on NovoCyte 3005 flow cytometer. Data was analyzed using FlowJo.

[0449] 2. HLA-G Off-Target Screen

[0450] HEK293T cells were transfected with plasmid encoding HLA-A, -B, -C, -E or -F using the Lipofectamine 3000. Two days after transfection, cells were harvested and stained sequentially with viability dye, primary antibody (mAb clone or human lgG1 isotype control) at 1.5 ug / ml, and then finally with anti-human IgG-APC at 5 ug / ml. A commercial antibody was used as a positive control to demonstrate the appropriate HLA expression on the HEK293T cell line after transfection. Cell suspensions were acquired on NovoCyte 3005 flow cytometer. Data was analyzed using FlowJo.

[0451] 3. Direct ELISA

[0452] ELISA plates were coated with 1 pg / mL human HLAG complex tetramer in PBS overnight at 5 oC. Then blocked with 3%BSA in PBST for 2-3 hours at room temperature. 3-fold serially diluted antibodies starting at 10 pg / mL were applied onto the plates and incubated for 1 h at room temperature. The human HLAG complex tetramer-bound antibodies were detected by HRP- conjugated rabbit anti human IgG-Fc at a 1 :10,000 dilution. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and EC50’s were calculated with GraphPad Prism 8.0 software.

[0453] 4. Competitive ELISA

[0454] ELISA plates were coated with 5 pg / mL LILRB2-Fc in PBS overnight at 5 oC. Then blocked with 3%BSA in PBST for 2-3 hours at room temperature. During the blocking step, each HLAG mAb was pre-incubated at 10pg / mL with 10, 3.33, and 1.11 pg / mL HLAG complex tetramer (HLAG-BT) for 1 hour at room temperature. After 1 hour the preincubate mAb was added with HLAG-Bt to the ELISA plate and incubated for 1 hour at room temperature. HRP-conjugated Streptavidin at a 1 :5,000 dilution was added and incubated for 1h at room temperature. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Percent blocking was calculated as 100 - (Absorbance of mAb + HLAGBT / absorbance of HLAG-Bt) *100.

[0455] 5. Expression and Purity

[0456] All clones were purified through Protein A affinity chromatograph followed by CHT chromatograph. Green Juice (GJ) expression was determined by ProA HPLC method (LAB-116). Product recovered after full purification measured by A280 (LAB-102). Purity was determined by SEC-HPLC (LAB- 108).

[0457] 6. Thermostability

[0458] Each samples was loaded into a Uni and ran on the Uncle (Unchained labs). The Uni was heated from 5 oC to 95 oC in 0.05 oC / s increments, and the Uncle reads three different measurement modes — fluorescence, Static Light Scattering (SLS) and Dynamic Light Scattering (DLS). Tm1 and Tm2 are measured from intrinsic fluorescence melt curves. Tagg 266 and Tagg473 were determined by SLS.

[0459] 7. JEG-3 - U937: Immunosuppression Assay

[0460] Human monocyte U937 cells (Sigma) were grown in suspension in complete 1x RPMI (10% FBS, 1x pen / strep, 1x glutamax) prior to differentiation. U937 cells were differentiated in complete media with a final concentration of 10 nM Phorbol 12-myristate 13-acetate (PMA) (Sigma) for 48 hours at 37°C. Human JEG-3 cells (ATCC) were plated in 96-well tissue treated plates at a density of 1.5e4 cells / well in 1x EMEM (10% FBS, 1x penicillin / streptomycin, 1x glutamax) until 90-100% confluent or 48 hours at 37°C. After 48 hours HLAG mAbs were diluted to 2x the desired concentration in complete 1x RPMI media. The 1x EMEM media was removed and 100 pL of HLAG mAbs were added to the JEG-3 96-well plate for a 1-hour pre-incubation. Following the pre-incubation 100 pL of differentiated U937 cells in complete 1x RPMI were added to the confluent JEG-3 96-well plate at a density of 5e5 cells / mL. The final concentration of the HLAG mAbs was 80, 20, and 5 pg / mL. The plate was incubated for 20 hours at 37°C. The next day LPS-EB (Invivogen) was reconstituted according to the manufacturer’s recommendation and added at a final concentration of 50 ng / mL in 1x RPMI to desired wells for 5 hours at 37°C. After 5 hours cell culture supernatants were retained for TNFa ELISAs performed using TNFa Human Uncoated ELISA kit (Thermo Fisher) in accordance with the manufacturer’s instructions. 8. Further in vitro and in vivo studies of KB17A5.3

[0461] To support the preclinical development of KB17A, several in vitro and in vivo evaluations have been performed which are summarized below and described in more detail in the following sections. In vitro experiments were first performed to confirm binding specificity of KB17A.5.3 for human HLA- G, then to measure the functional activity of KB17A.5.3 as an HLA-G antagonist blocking receptor interaction with LILRB2. Follow-on experiments were performed to define the epitope specificity and screen for cross-reactivity between KB17A.5.3 and HLA-receptor family homologs. Cell culture experiments performed using both commercially-available assay kits as well as primary human effector cells demonstrated that treatment of human immune cells with KB17A.5.3 prevents HLA-G induced immune suppression of inflammatory cytokine production following exposure to an inflammatory stimulus.

[0462] In vivo efficacy of KB17A.5.3 has been demonstrated in two humanized (CD34+ HuHSC-NOG-EXL) murine xenograft studies. In both studies, HLA-G expressing melanoma or bladder carcinoma cells were injected subcutaneously into mice which was followed by KB17A.5.3 treatment. The studies show that KB17A.5.3 treatment significantly reduces tumor growth and induces mechanism-driven immune cell activation in tumors of humanized mice.

[0463] The completed and planned nonclinical studies of KB17A.5.3 included herein are summarized in

[0464] Table 5 below.

[0465] Table 5 Nonclinical Studies of KB17A.5.3

[0466] 8.1 In Vitro Studies of KB17A.5.3

[0467] A panel of in vitro methods was used to define the binding affinity and specificity of the KB17A.5.3 CDR for human HLA-G, as well as to measure KB17A Fc binding to Fc receptors relevant to immune cell signaling. These methods included ELISAs using human HLA-G complex tetramer (Kactus and Reprokine both with accession number P17693), Cynomolgus HLA-G tetramer protein (Kactus, HLG-CM41CT) or Rhesus macaque HLA-G tetramer protein (Kactus, HLG-RM41CT), as well as flow cytometry and surface plasmon resonance (SPR) experiments focusing on binding activity of the KB17A.5.3 CDR. A competitive ELISA was also used to confirm that KB17A.5.3 blocks the binding of recombinant HLA-G to recombinant LILRB2-Fc. Finally, a panel of ELISA methods was used to assess interactions between the KB17A.5.3 Fc and a variety of human Fc receptors important for macrophage and NK cell mediated ADCC activity including CD64, CD32a, CD32b / c and CD16a. These experiments demonstrated that the KB17A.5.3 CDR binds strongly and specifically to human HLA-G with no cross-reactivity to human HLA-receptor family homologs and no observable cross-reactivity for non-human primate HLA-G; that KB17A.5.3 binding to HLA-G prevents binding interactions between HLA-G and LILRB2; and that the KB17A.5.3 Fc is fully functional and binds with Fc receptors relevant to immune signaling.

[0468] Mechanistic evaluations of KB17A.5.3 included several in vitro tissue culture experiments designed to measure the extent to which KB17A.5.3-mediated HLA-G antagonism reverses suppression of human monocytes under conditions representative of a tumor microenvironment (i.e., in the presence of both HLA-G-expressing cells and an inflammatory stimulus), as well as to measure KB17A.5.3-mediated antibody-dependent cellular cytotoxicity (ADCC) in the presence of human primary immune effector cells. These experiments demonstrated that KB17A.5.3 treatment of HLA- G-suppressed monocytes restores immune reactivity to inflammatory stimuli, and that KB17A.5.3 binding to HLA-G expressing cells elicits ADCC transcriptional activity in Jurkat cells as well as direct cytotoxicity in the presence of human peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells.

[0469] A sequence-identical version of the TTX-080 anti-HLA-G mAb was produced in the Nb7KOAXylT / FucT plant host (Nicotiana benthamiana) and used as a control / comparator antibody in select in vitro experiments, as identified in the sections below.

[0470] 8.1.1 Human HLA-G Binding

[0471] To confirm KB17A.5.3 binding to human HLA-G, ELISA plates were coated with 1 pg / mL human HLA-G complex tetramer in PBS overnight at 5 °C then blocked with 3% BSA in PBST for 2-3 hours at room temperature. 3-fold serial dilutions of KB17A.5.3 starting at 10 pg / mL were applied onto the plates and incubated for 1 h at room temperature. The human HLA-G complex tetramer-bound antibody was detected by HRP-conjugated rabbit anti human IgG-Fc at a 1 :10,000 dilution. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and EC50’s were calculated with GraphPad Prism 8.0 software. Antibody TTX-080 produced in Nicotiana benthamiana was used as a positive binding control. Results are shown in Figure 9 and demonstrate that KB17A.5.3 binds to human HLA-G with an EC50 of 16 ng / mL. 8.1.2 Non-Human Primate HLA-G Binding

[0472] To assess KB17A.5.3 cross-reactivity to non-human primate (NHP) HLA-G orthologs in support of nonclinical safety species selection, a literature review of HLA-G homology was performed. Published functional and sequence comparisons of human HLA-G with the closest known canine (DLA) and murine (Qa-2) homologs do not support the relevance of either species for nonclinical safety assessments of KB17A.5.3 (Clements 2005, Yuhki 2007, Da Silva 2018), nor are suitable reagents available for canine or murine KB17A.5.3 binding assessments due to the absence of a known HLA-G ortholog(s) in either species. The only well-characterized nonclinical safety species with a known HLA-G ortholog is NHP, and reagents were available for both Rhesus and Cynomolgous macaques to perform an ELISA-based analysis of KB17A.5.3 cross reactivity for endogenous HLA-G in these two primate models.

[0473] ELISA plates were coated with 2.5 pg / mL of either KB17A.5.3, plant-made TTX-80 positive control, and plant-made I gG 1 isotype control in PBS overnight at 5 °C, then blocked with 5% BSA in PBST for 3-4 hours at room temperature. 3-fold serial dilutions starting at 30 pg / mL were applied onto the plates of Cynomolgus (Kactus, H LG- CM 41 CT), Rhesus macaque (Kactus, HLG-RM41CT), and Human (Reprokine, MHCHLA-GT) HLA-G tetramer protein and incubated for 1 hour at room temperature. Bound protein was detected by HRP-conjugated mouse anti human his tag at a 1 :5,000 dilution. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and EC50’s (pg / mL) were calculated with GraphPad Prism 8.0 software.

[0474] Results are shown in Figure 10 and show that KB17A.5.3 did not demonstrate binding affinity for either Cynomolgus (upper graph) or Rhesus macaque (lower graph) HLA-G but bound with high affinity to human HLA-G.

[0475] 8.1.3 Binding Confirmation by Flow Cytometry

[0476] Flow cytometry was used to confirm KB17A.5.3 binding to cell surface human HLA-G and to measure cross-reactivity to HLA-family homologs. HEK293T cells were transfected with plasmids expressing HLA-G or homologs HLA-A, HLA-B, HLA-C and HLA-E. After 2 days to allow for gene expression, cells were stained with KB17A.5.3 (red histogram), isotype control (grey filled histogram), or a commercial antibody against corresponding protein (black dotted histogram). On- target HLA-G staining was performed using 0.15 pg / ml of KB17A.5.3 or isotype control. Off-target homolog staining was performed using 1.5 pg / ml of KB17A.5.3 or isotype control. The dose curve was serial dilutions of KB17A.5.3 or isotype starting at 10 pg / mL. Geometric mean fluorescence intensity (gMFI) of KB17A.5.3 was compared to the gMFI of the isotype control.

[0477] Results are shown in Figure 11 and KB17A.5.3 binds to HLA-G overexpressing HEK293T cells down to a concentration of 0.0015 pg / mL; at 1.5 pg / mL, KB17A.5.3 does not bind to HLA -A, -B, -C, or - E. HEK293T cells transfected with HLA-F plasmid was tested in ELISA format and binding was not found. Thus, KB17A.5.3 has strong cell-based binding to HLA-G with no significant cross reactivity to other HLA family members.

[0478] Further results are provided in Figure 12 demonstrating KB17A.5.3 binding for the endogenous HLA- G expressing cell-line, JEG-3. KB17A.5.3 binds to JEG-3 cells, with an EC50 of 78.6 ng / mL. Graphs were plotted and EC50’s were calculated with GraphPad Prism 8.0 software.

[0479] 8.1.4 Binding Confirmation by Surface Plasmon Resonance

[0480] The binding affinity (KD) of KB17A.5.3 to biotinylated HLA-G (BT-HLA-G) was measured on a Biacore T200 instrument at ambient temperature. BT-HLA-G was captured on an SA sensor chip following the manufacturer’s instructions to a surface density of about 200 RUs. Serial dilutions of HLA-G mAb were made in running buffer (HBS-EP+, Cytiva) and injected at a flow rate of 30 pL / min for a contact time of 60s and a dissociation time of 600s. Between sample injections, the system was washed with wash buffer. A reference flow cell was utilized to correct response contributions such as bulk shifts that occur equally in the sample and reference flow cells. A blank cycle (running buffer) was performed, and all sample injections were blank subtracted to correct the sensorgrams for drifts and other disturbances that affect the reference subtracted curve. A replicate of a non-zero concentration of mAb and the blank were injected in each experiment for double referencing, thus verifying the reliability of the SA chip throughout the experiment. The data were assessed by kinetics 1 :1 binding analysis for Association rate constant (ka), Dissociation rate constant (kd), and Equilibrium dissociation constant (KD). The data is presented in Table 6 below:

[0481] Table 6: Binding Confirmation of KB17A.5.3 by SPR Results are shown in Figure 13 which demonstrates KB17A.5.3 binds to HLA-G with high affinity and corroborates the ELISA and flow cytometry binding data. The positive control was TTX-080 produced in Nicotiana benthamiana. SPR was performed in triplicate and error bars represent standard deviation.

[0482] 8.1.5 KB17A.5.3 LILRB2 / HLA-G Blocking by ELISA

[0483] Additional ELISA experiments were performed to confirm that KB17A.5.3 blocks the binding of recombinant HLA-G to recombinant LILRB2-Fc. ELISA plates were coated with 5 pg / mL LILRB2-Fc in PBS overnight at 5°C, then blocked with 3% BSA in PBST for 2-3 hours at room temperature. During the blocking step, a KB17A.5.3 dose curve starting at 100 pg / mL (3-fold dilutions) was preincubated with 1 pg / mL HLA-G complex tetramer (HLA-G-BT) for 1 hour at room temperature. After 1 hour the preincubated mAb with HLA-G-BT was added to the ELISA plate and incubated for 1 hour at room temperature. HRP-conjugated Streptavidin at a 1 :2,500 dilution was added and incubated for 1h at room temperature. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader and IC50 was calculated in GraphPad Prism.

[0484] Results are shown in Figure 14 demonstrating that KB17A.5.3 exhibited blocking activity with an IC50 of 0.338 pg / mL confirming that the antibody is able to block binding association between recombinant HLA-G and LILRB2. TTX-080 produced in Nicotiana benthamiana was used as a positive control and has an IC50 of 0.268 pg / mL.

[0485] 8.1.6 Fc Receptor Binding by ELISA KB17A.5.3 binding to human Fc receptors that are important for macrophage and NK cell mediated ADCC activity was investigated using ELISA assays. To confirm KB17A.5.3 binding to human Fc receptors CD64 and CD 16a, ELISA plates were coated with 1 pg / mL human Fc receptor [(CD64, FCA-H52H1); (CD16a, CD8-H52H4); Aero Biosystems] antigens in PBS overnight at 5°C then blocked with 5% BSA in PBST for 3-4 hours at room temperature. 3-fold serial dilutions of KB17A.5.3 starting at 10 pg / mL were applied onto the plates and incubated for 1 hour at room temperature. The human Fc receptor-bound antibody was detected by HRP-conjugated rabbit anti human IgG- Fc at a 1 :10,000 dilution. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and EC50’s were calculated with GraphPad Prism 8.0 software. These ELISAs were done three times to show an average and standard deviation, and a representative graph is shown.

[0486] For CD32a and CD32b / c, ELISA plates were coated with 2.5 pg / mL human Fc receptor [(CD32a, CDA-H5221); (CD32b / c, CDB-H5228); Aero Biosystems] antigens in PBS overnight at 5°C then blocked with 5% BSA in PBST for 3-4 hours at room temperature. 2-fold serial dilutions of KB17A.5.3 starting at 3 mg / mL were applied onto the plates and incubated for 1 hour at room temperature. The human Fc receptor-bound antibody was detected by HRP-conjugated rabbit anti human IgG-Fc at a 1 :10,000 dilution. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and EC50’s were calculated with GraphPad Prism 8.0 software. These ELISAs were done one time.

[0487] Results are shown in Figure 15 and demonstrate that KB17A.5.3 binds strongly to human Fc receptors CD64 (EC50, 38.61 ± 17.8 ng / mL) and CD16a (EC50, 108.05 ± 91.9 ng / mL) relative to positive control (commercially-obtained Humira). Additionally, KB17A.5.3 binds to CD32b / c (EC50, 171.5 pg / mL) and CD32a (EC50, 6.5 pg / mL).

[0488] 8.1.6 FcRn Binding by ELISA To confirm KB17A.5.3 binding to human FcRn, ELISA plates were coated with 2.5 pg / mL human FcRn antigen (Aero Biosystems, FCN-H52W7) in PBS overnight at 5°C then blocked with 5% BSA in PBST for 3-4 hours at room temperature. 2-fold serial dilutions of KB17A.5.3 starting at 3 mg / mL were applied onto the plates and incubated for 1 hour at room temperature. The human FcRn receptor-bound antibody was detected by HRP-conjugated rabbit anti human IgG-Fc at a 1 :10,000 dilution. The tetramethylbenzidine (TMB) substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and ECso’s were calculated with GraphPad Prism 8.0 software.

[0489] Results are shown in Figure 16, demonstrating that KB17A.5.3 binds to neonatal Fc Receptor with an ECso of 170.8 pg / mL.

[0490] 8.1.7 Prevention of HLA-G Induced Monocyte Suppression

[0491] To measure KB17A.5.3’s ability to prevent HLA-G induced monocyte suppression, human monocytes were co-cultured with KB17A.5.3 in a cell-based assay as follows. Cryopreserved CD14 single donor human monocyte cells (PromoCell & Stem Cell Technologies) were cultured in mononuclear cell media (PromoCell) for 18-24 hours prior to the assay. Human JEG-3 cells or HT- 1376 cells (ATCC) were plated in 96-well tissue treated plates at a density of 1.5e4 cells / well in complete EMEM until 90-100% confluent or 48 hours at 37°C. After 48 hours the EMEM media was removed, and 100 pL of KB17A.5.3 was added at a final concentration of 80, 5, 1.25, or 0.3125 pg / mL for 1 hour at 37°C. CD14 monocytes were transferred to complete RPMI media and added to the JEG-3 and mAb mixture at a density of 7.5e4 cells / well. The plate was incubated for 20 hours at 37°C. The next day LPS-EB (Invivogen) was reconstituted according to the manufacturer’s recommendation and added at a final concentration of 100 ng / mL in RPMI media to desired wells for 6 hours at 37°C. After 6 hours cell culture supernatants were retained for TNF-a ELISAs performed using TNF-a Human Uncoated ELISA kit (Thermo Fisher) in accordance with the manufacturer’s instructions.

[0492] Results are shown in Figure 17 which demonstrates that KB17A.5.3 prevents HLA-G induced monocyte suppression of TNF-a cytokine release in a dose-dependent manner when compared to controls in four donors. Figure 17 is representative of one donor. Treatment with plant-made lgG1 isotype control results in no prevention of HLA-G mediated suppression of TNF-a. Percent TNF-a release by KB17A was calculated between treatment groups and stimulation control. The # indicates > 80% TNF-a release was maintained in comparison to Stimulation control. 8.1.8 Effector Functional Activity (ADCC) Using Primary Cells

[0493] A cell-based assay was developed to assess KB17A.5.3-mediated antibody-dependent cellular cytotoxicity (ADCC) from human peripheral blood mononuclear cells (PBMCs) when bound to human HLA-G-expressing cells. Human PBMCs from 2 separate donors (PromoCell & Stem Cell Technologies) were cultured at 37°C in mononuclear cell media (PromoCell) 18-24 hours prior to the assay. The human choriocarcinoma cell line JEG-3 (ATCC HTB-36) endogenously expresses HLA-G and was maintained in 1x EMEM complete media prior to the assay. At the start of the assay, JEG-3 target cells were labeled with 0.25% BATDA dye (Revvity, C136-100) at 37 °C for 20 minutes, which can unidirectionally penetrate cells, and is released upon lysis. The released dye reacts with Europium forming a fluorescent chelate (EuTDA), which can be measured. The BATDA labeled JEG-3 cells were then washed via centrifugation at 250 x g for 2 minutes and resuspended in 10% FBS 1x PBS solution six times. On the last wash the labeled JEG-3 cells were resuspended in 1x RPMI complete media. KB17A.5.3 and control mAbs were diluted at optimized dose ranges for the individual PBMC donors. PBMCs were added at an E:T ratio of 50:1 to labeled JEG-3 target cells at 5e3 cells / well in a 96-well plate and incubated at 37°C for 2.5 hours. Cell mixture supernatants were added at a 1 :10 ratio with Europium solution and incubated at room temperature for 15 minutes. Time-resolved fluorescence was utilized, the emission was measured at 615 nm, and excited at 337 nm. Calculations were conducted using the Revvity DELFI EuTDA Cytotoxicity Detection kit (AD0116) manufacturer’s instructions.

[0494] Results are shown in Figure 18 which demonstrates that KB17A.5.3 elicited cytolysis of HLA-G expressing JEG-3 when in co-culture with primary human PBMC cells. In contrast, plant-made I gG 1 isotype control and TTX-080 produced in Nicotiana benthamiana (termed ‘positive control null’ because the antibody lacks an Fc) did not demonstrate any cytolysis above spontaneous release. KB17A.5.3 demonstrated ADCC activity with PBMC cells in multiple donors (Donor 1 : EC50, 0.052 pg / mL) (Donor 2: EC50, 0.023 pg / mL) and is effective at targeting HLA-G expressing cells.

[0495] 8.1.9 Effector Functional Activity (ADCC) Using Natural Killer (NK) Cells KB17A.5.3 was separately assessed for its ability to stimulate an ADCC response from NK cells using flow cytometry. Natural Killer (NK) cells were isolated from fresh human peripheral blood (leukopheresis) using negative selection with magnetic beads. NK cells were then co-cultured with CellTrace Violet-labelled (HLA-G+) JEG-3 cells in the presence of KB17A.5.3 or isotype control (both at 10 pg / mL) at various effector-to-target ratios. After 4-hour incubation, JEG-3 viability was assessed by flow cytometry. The data was normalized to account for spontaneous non-specific death of the target cells.

[0496] Results are shown Figure 19 which demonstrates that when primary human NK cells were cocultured with HLA-G-expressing JEG-3 cells, KB17A.5.3 elicited robust cytotoxicity of JEG-3 cells. In contrast, a plant-made lgG1 isotype control did not demonstrate any killing above background. KB17A.5.3 therefore is very effective at targeting HLA-G-expressing cells for killing by NK cells.

[0497] 8.1.10 Effector Functional Activity (ADCC) Using Jurkat Cells

[0498] KB17A.5.3 was also assessed for ADCC activity using a reporter gene assay with Jurkat effector cells that stably express the FcyRllla receptor and NFAT response element. Luciferase is produced from NFAT pathway activation via concomitant binding of target and effector cells. Promega’s ADCC Reporter Bioassay kit (G7015) was utilized to assess KB17A.5.3-mediated ADCC activity in Jurkat cells co-incubated with JEG-3 (HLA-G+) cells. The human choriocarcinoma cell line JEG-3 (ATCC HTB-36) endogenously expresses HLA-G and was maintained in 1x EMEM complete media prior to the assay. JEG-3 (HLA-G+) target cells were seeded at 1.5e4 cells / well in a 96-well plate for 48 hours at 37°C. After 48 hours, 4% low IgG serum was diluted in RPMI to prepare KB17A.5.3 dilutions and ADCC effector cells. KB17A.5.3 (in duplicate) was diluted for a final concentration of 10 ug / mL with five 10-fold serial dilutions. EMEM media was removed from JEG-3 target cells and then KB17A.5.3 was added. This mixture was incubated at 37°C for 30 minutes. After the incubation the ADCC effector cells were added to target cells at 7.5e4 cells / well for 6 and 23 hours at 37°C. Bio- Glo Luciferase Assay Reagent was added to desired wells at a 1 :1 ratio with the cell mixture. Luminescence was measured and calculations were performed according to the kit manufacturer’s instructions.

[0499] Results are shown in Figure 20, which demonstrates that KB17A.5.3 elicited ADCC reporter gene activity in the assay after incubation periods of 6 hours (EC50, 0.064 pg / mL) and 23 hours (EC50, 0.070 pg / mL).

[0500] 8.2 In Vivo Studies of KB17A.5.3 KB17A.5.3 has demonstrated anti-tumor activity in two humanized mouse xenograft models of HLA- G expressing melanoma or bladder carcinoma.

[0501] In the first proof-of-concept in vivo study, KB17A.5.3 was tested in a humanized mouse model (NOG-EXL mice reconstituted with human hematopoietic cells (HuHSC)) wherein mice were inoculated with HLA-G overexpressing SK-MEL-5 melanoma cells and tumor growth was monitored until average tumor size reached 100 mm3(Day 0). Randomized groups of mice, each containing mice from three different HuHSC donors, were intravenously (I.V.) dosed with isotype control antibody (hlgG1 20 mg / kg) or KB17A.5.3 (hlgG1 20 mg / kg) twice a week for 31 days. Tumor volume was monitored until Day 38. The results indicate significant tumor growth inhibition (-30%) starting 21 days after treatment in mice treated with KB17A.5.3 compared to animals dosed with isotype control. Additionally, KB17A.5.3 treatment increased the number of CD8+ T cells and the percentage of CD11 b+CD33+ cells in the tumor microenvironment compared to isotype control treated animals.

[0502] In a second confirmatory xenograft study, HuHSC-NOG-EXL mice were inoculated with endogenous HLA-G and PD-L1 expressing HT-1376 (bladder carcinoma) cells and tumor volume was monitored until average tumor sizes reached 100 mm3(Day 0). Randomized groups of mice, each containing mice from three different huHSC donors, were intraperitoneally dosed with isotype control antibody (hlgG1 10 mg / kg), KB17A.5.3 (hlgG1 10 mg / kg), Atezolizumab (hlgG1 10mg / kg), or KB17A.5.3 and Atezolizumab in combination (10 mg / kg each) twice a week starting on Day 1 for five total doses. Tumor volume was monitored until Day 17. The data indicates impaired tumor growth over the course of the study in mice treated with KB17A.5.3 or Atezolizumab as demonstrated by significant reduction in tumor growth and increased CD8+ T-cells and CD33+CD11b-CD86+ cell subsets in the tumor microenvironment. KB17A.5.3 treatment reduced tumor volume by -30% compared to isotype control, similar to atezolizumab positive control treatment after 17 days.

[0503] A Winn assay mouse model of KB17A.5.3 treatment using freshly isolated human PBMCs and JEG- 3 placental cancer cells is being developed. The final planned Winn study will include a dose-ranging phase to support dose justification for the first-in-human study of KB17A.5.3.

[0504] 8.2.1 Proof-Of-Concept Efficacy in Humanized Mouse Xenograft Model

[0505] Study Design A preliminary proof-of-concept in vivo efficacy study was performed to assess KB17A.5.3 in the treatment of subcutaneous HLA-G overexpressing SK-MEL-5 melanoma in a xenograft model in female HuHSC-NOG EXL mice. A human I gG 1 isotype antibody produced by KBio (EVD68-228-N) was used as a negative control. In this model, immunodeficient NOG-EXL (NOD.Cg- Prkdcscidll2rgtm1SugTg(SV40 / HTLV-IL3,CSF2)10-7Jic / JicCrl)) mice were irradiated and injected with human CD34+ hematopoietic stem cells (HSC) isolated from umbilical cord blood of three separate donors. Eight to twelve weeks post CD34+ HSC transplant, >40% human CD45+ immune cells were detected in mouse blood. The mice were then implanted subcutaneously (SC) with 5 x 106human melanoma derived HLA-G overexpressing SK-MEL-5 cell line. HuHSC-NOG-EXL mice bearing SK- MEL-5 tumors were assigned to treatment groups at 9 mice per group (3 mice from each human CD34+ donor cohort per treatment group) when mean tumor size was 100 mm3after approximately 32 days, (Day 0 tumor randomization). Tumor-bearing mice were injected I.V. with 20 mg / kg of KB17A.5.3 or a hlgG1 isotype control mAb twice a week for 31 days (10 total doses) following randomization. Tumor volumes were monitored twice weekly following the initiation of treatment.

[0506] Table 7 Proof-of-Concept Mouse Xenograft Efficacy Study Design

[0507] 1Animals were dosed on Days 0, 3, 7, 10, 14, 17, 21 , 24, 28, and 31 (10 total doses).

[0508] Tumor Cell Preparation

[0509] HLA-G overexpressing SK-MEL-5 cells were maintained in vitro in MEM medium supplemented with 10% fetal bovine serum, 0.01 mM NEAA at 37°C in an atmosphere of 5% CO2 in air. Cells were seeded by adding 1.49 x 106cells per T25 tissue culture flask and propagated until cells exceeded a density of 80%. Subsequently, cells were harvested using trypsinization and subcultured at a 1 :2- 1 :4 ratio. The cells in exponential growth phase (a density of >80%) were harvested and quantitated by cell counter before tumor inoculation.

[0510] Tumor Implantation Each mouse was inoculated subcutaneously in the right upper flank region with HLA-G overexpressing SK-MEL-5 cells (5 x 106) in 0.2 mL of PBS mixed with Matrigel (1 :1) for tumor development. Randomization started when the mean tumor size reached 99.44 mm3. The date of randomization was denoted as day 0; treatment was initiated on the same day of randomization (day 0).

[0511] Tumor Measurement

[0512] Tumor volumes were measured twice per week after randomization in two dimensions using a caliper, and the volume was expressed in mm3using the formula: “V = (L x Wx W) / 2, where V was tumor volume, L was tumor length (the longest tumor dimension), and W was tumor width (the longest tumor dimension perpendicular to L). Dosing as well as tumor and body weight measurements were conducted in a Laminar Flow Cabinet.

[0513] After tumor cell inoculation, animals were checked daily for morbidity and mortality. During routine monitoring, the animals were checked for any effects of tumor growth and treatments on behavior such as mobility, food and water consumption, body weight gain / loss (Body weights were measured twice per week after randomization), eye / hair matting and any other abnormalities. Mortality and observed clinical signs were recorded for individual animals in detail.

[0514] The body weights of all animals were monitored throughout the study and animals were euthanized if they lost over 20% of their body weight relative to the weight on the day of randomization. Mice were euthanized if tumor volume exceeded 3000 mm3. To deter cannibalization, any animals exhibiting an ulcerated or necrotic tumor were separated immediately and singly housed and monitored daily before the animal was euthanized or until tumor regression was completed. Mice were euthanized rapidly if a) tumors ulcerated, and the ulceration diameter was greater than 5 mm, or pus or necrosis observed; or b) tumor burden, including metastasis, compromised animal’s normal physiologic performances, e.g., orientation, access to food or water, etc. Mice were also euthanized rapidly for any of the following reasons:

[0515] Emaciated - skeletal structure extremely prominent, little or no flesh cover, vertebrae distinctly segmented.

[0516] Persistent hypothermia.

[0517] Blood stained or mucopurulent discharge from any orifice.

[0518] Labored breathing, particularly accompanied by nasal discharge or cyanosis.

[0519] Hind limb paralysis or weakness (cannot reach food and water).

[0520] Severe anemia, mainly indicated by pale feet or ear.

[0521] Severe infection.

[0522] Incontinence or diarrhea over 48 hours or severe dehydration. Randomization

[0523] The randomization started when the mean tumor size reached approximately 100 mm3. All animals were randomly allocated to study groups, 9 mice in each group. Randomization was performed based on “Matched distribution” method.

[0524] The date of randomization was denoted as day 0.

[0525] Study Termination

[0526] The study was terminated on day 38.

[0527] Test and Control Articles

[0528] Table 8 Test and Control Articles

[0529] 8.2.2 Tumor Volume Growth

[0530] The treatment (20 mg / kg) was performed twice a week (Bl W) for 31 days. T umor volumes were also measured twice per week after randomization until termination on Day 38. Day 0 started when the mean tumor size reached 99.44 mm3and mice were randomized into groups with 3 mice per HSC CD34+ donor (n=9). Of the three cohorts of CD34+ engrafted mice in the Isotype control group, one cohort showed inconsistent tumor growth and has been removed from the analysis shown below. As shown in Figure 21 , the mean change in tumor volume from Day 0 was significantly reduced starting on Day 21 in KB17A.5.3 treated animals compared to Isotype control. On Day 38, the mean tumor volume in the KB17A.5.3 treatment group was 32.8% (271.4 mm3) less than Isotype control group (Table 9).

[0531] 8.2.3 Tumor Growth Inhibition

[0532] T umor Growth Inhibition (TGI) and T umor volume of the treatment control groups (T / C) calculation based on the tumor size data from Day 38 are presented in Table 9.

[0533] Table 9 TGI and T / C calculation on Day 38

[0534] Tumor Growth Inhibition; TGI% = (1-Ti / Ci) x 100. The Ti / Ci value (%): T / C% = Ti / Ci x 100%. Ti and Ci are the mean tumor volumes of the treatment and control groups, respectively, on a given day.

[0535] 8.2.4 Tumor Weights

[0536] At the end of study on Day 38, tumors were collected and weighed. The mean tumor weights are plotted in Figure 22.

[0537] 8.2.5 Flow cytometry analysis of tumor immune cells

[0538] At termination, all tumor samples were processed as single cell suspensions and stained for the following panel of markers: Live / Dead Dye, mCD45, hCD45, hCD3, hCD4, hCD8, hHLA-DR, hCD33, hCD11 b, hCD86 with absolute count according to the protocol, followed by flow cytometry data acquisition and analysis. The final statistical analyses were performed on proportions of cells of interest from all cells of the population.

[0539] Results are shown in Figure 23.

[0540] 8.2.6 Flow cytometry analysis of Human Blood CD45+ Immune Cells

[0541] At termination, all blood samples were stained for the following panel of markers: Live / Dead Dye, mCD45, hCD45 according to the protocol, followed by flow cytometry data acquisition and analysis. The final statistical analyses were performed on proportions of cells of interest from all cells of the population.

[0542] Results are shown in Figure 24.

[0543] 8.2.7 Analysis of HLA-G Protein Expression in Tumors by Western Blot

[0544] Tumor samples were harvested on Day 38 and analyzed via Western Blot assay to determine if HLA-G expression was maintained in all tumors. As presented in Figure 25, hHLA-G was expressed in all tumors.

[0545] 8.2.8 Conclusion - Proof-Of-Concept Efficacy in Humanized Mouse Xenograft Model

[0546] In this first proof-of-concept in vivo study, the therapeutic efficacy of KB17A.5.3 was investigated in a SK-MEL-5 (melanoma) cell line derived subcutaneous xenograft model in HuHSC-NOG-EXL mice.

[0547] Significant anti-tumor effects were observed in KB17A.5.3 treated HuHSC-NOG-EXL mice engrafted with human cord blood-derived CD34+ cells from two separate donors. Significant antitumor effects were detected in KB17A.5.3 treated animals starting on Day 21 compared to hlgG1 Isotype Control group. The TGI compared to the hlgG1 Isotype Control group was 32.80% in KB17A.5.3 treated animals.

[0548] Flow cytometry analysis of tumors on Day 38 revealed neither treatment with Isotype control nor KB17A.5.3 had significant impacts on the human CD45+ immune cell reconstruction rate in blood. In contrast, in the tumors of KB17A.5.3 treated animals, significant increases in the absolute number of CD3+ and CD8+ T cells were observed. For myeloid cells, KB17A.5.3 treatment resulted in significantly increased percentages of CD11b+CD33+ subsets.

[0549] In conclusion, this study established that HLA-G overexpressing SK-MEL-5 tumors grow in CD34+ NOG-EXL mice and maintain HLA-G expression throughout the study. Furthermore, significant evidence of anti-tumor activity was observed in this study with KB17A.5.3 as demonstrated by significant reduction in tumor growth and increased immune cells in the tumor microenvironment.

[0550] 8.2.9 Confirmatory Efficacy in Humanized Mouse Xenograft Model

[0551] Study Design A follow-on confirmatory in vivo efficacy study was performed to assess KB17A.5.3 in the treatment of subcutaneous HLA-G expressing HT-1376 bladder cancer cells in a xenograft model in female HuHSC-NOG EXL mice (Table 11). A human IgG 1 isotype antibody produced by KBio (EVD68-228- N) was used as a negative control. A commercial anti-PDL1 antibody (Tecentriq; Atezolizumab) was procured and used as a positive control.

[0552] Table 11 Confirmatory Mouse Xenograft Efficacy Study Design

[0553] 1Animals were dosed on Days 1 , 4, 8, 11 , and 15.

[0554] Tumor Cell Preparation

[0555] Cryogenic vials containing tumor cells were thawed and cultured according to the manufacturer’s protocol. On the day of injection, cells were washed in serum-free media, counted, and resuspended in cold serum-free media at a concentration of 10M viable cells / 100 pL.

[0556] Cell suspensions were mixed with an equal volume of Matrigel ECM resulting in a final concentration of 5M viable cells / 100 pL. The ECM-cell mixture was kept on ice during transport to the vivarium. Cells were prepared for injection by withdrawing the ECM-cell mixture into a chilled 1 mL slip-tip syringe. The filled syringes were kept on ice to avoid solidification of the ECM.

[0557] Tumor Implantation

[0558] Animals were shaved and ear tagged prior to injection. One mouse at a time was immobilized and the site of injection was disinfected with an alcohol swab. 100 pL of the cell suspension was subcutaneously injected into the rear flank of the mouse. Up to 5 animals were injected with 100 pL of cell suspension per syringe. Animals were undisturbed for up to seven days before observing for tumor growth.

[0559] Tumor Measurement Animals were monitored weekly for palpable tumors, or any changes in appearance or behavior. Once tumors were palpable, tumors were measured at least once a week using calipers. Tumor volumes were calculated using the following equation as described previously: (L*W*W) / 2. Once tumors were of appropriate size to begin the study (100mm3), tumors and body weights were measured at least 2 times per week for the duration of the study. One individual was responsible for tumor measurements for the duration of the study.

[0560] Randomization and Dose Selection

[0561] When the average tumor volume reached 100 mm3, mice were randomly assigned to the respective treatment groups and dosed within 24 hours of randomization.

[0562] Reference Day: Day 0 is defined by the day of randomization.

[0563] Study Termination

[0564] The study was terminated 17 days after randomization (Day 17).

[0565] Test and Control Articles

[0566] Table 12 Test and Control Articles

[0567] Results

[0568] HuHSC-NOG EXL mice with human cord blood-derived CD34+ cells from three separate donors were enrolled in the study. All animals were confirmed >25% hCD45+ cells in peripheral blood prior to enrollment. Randomization was performed in the Study Log software on Day 0 once tumors reached >100 mm3. The mean tumor volume for each group +SEM at randomization was as follows (Table 13): Table 13 Average Tumor Volume at Randomization (Day 0)

[0569] 8.2.10 Tumor Volume

[0570] The tumor volume growth curves between randomization grouping and study termination are shown in Figure 26, Figure 27, and Table 14 below.

[0571] In Figure 26, the treatment (10 mg / kg) was performed twice weekly (BIW) for 14 days (5 total doses) once tumor reached >100 mm3 (Day 0). The study was terminated on Day 17. The mean (SEM) tumor volume of each treatment group is plotted (n=9). A 2-way ANOVA with a Dunnett’s multiple comparison test) was performed to compare treatment groups to Isotype Control. P values are presented as * color coordinated with the group that is being compared to Isotype control;*P < 0.05, **P < 0.01 , ***P < 0.001.

[0572] In Figure 27, the mean ±SEM A tumor volume during the dosing period (Days 0-17) of each treatment group is plotted (n=9). The treatment (10 mg / kg) was performed BIW for 14 days (5 total doses) once tumor reached >100 mm3 (Day 0). The study was terminated on Day 17. A 2-way ANOVA with a Dunnett’s multiple comparison test) was performed to compare treatment groups to Isotype Control. P values are presented as * color coordinated with the group that is being compared to Isotype control; *P < 0.05, **P < 0.01 , ***P < 0.001.

[0573] Table 14 TGI and T / C calculation on Day 17

[0574] Tumor Growth Inhibition; TGI (%) = (1-Ti / Ci) x 100; Ti and Ci are the mean tumor volumes of the treatment and control groups, respectively, on a given day.

[0575] 8.2.11 Flow cytometry analysis of tumor immune cells

[0576] At termination, all tumor samples were processed as single cell suspensions and stained for the following panel of markers: Live / Dead Dye, mCD45, hCD45, hCD19, hCD56, hCD3, hCD4, hCD8, hH LA-DR, hCD33, hCD11 b, hCD86, hPD1 , and hCD69 with absolute count according to the protocol, followed by flow cytometry data acquisition and analysis. The final statistical analyses were performed on proportions of cells of interest from all cells of the population.

[0577] Results are shown in Figures 28 and 29.

[0578] 8.2.13 Conclusion

[0579] In this study, the therapeutic efficacy of KB17A.5.3 was investigated in the treatment of HT-1376 (human bladder carcinoma) cell line derived subcutaneous xenograft model in female huHSC-NOG EXL mice. Prior to study start, the expression of HLA-G and PD-L1 was confirmed on the HT-1376 cells via FLOW cytometry (data on file).

[0580] Significant anti-tumor effects were detected in the all-treatment groups within the HuHSC-NOG EXL mice with human cord blood-derived CD34+ cells from three separate donors (n=9). On Day 17 in treatment groups KB17A.5.3 and Atezolizumab, the TGI compared to the hlgG1 isotype control group was 31.71% and 27.18%, respectively. Among all groups, maximum TGI was observed on Day 10 in KB17A.5.3 treated mice with a TGI compared to the hlgG1 isotype control group of 50.95%.

[0581] In conclusion, this study established that HLA-G expressing HT-1376 tumors grow in CD34+ NOG- EXL mice. Significant evidence of anti-tumor activity was observed in this study with KB17A or Atezolizumab treatment as demonstrated by significant reduction in tumor growth and increased immune cells in the tumor microenvironment.

Claims

CLAIMS1. An antibody, or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising:(a) a CDRH1 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 37;(b) a CDRH2 selected from the group consisting of SEQ ID NO 38 to SEQ ID NO 77; and(c) a CDRH3 selected from the group consisting of SEQ ID NO 78 to SEQ ID NO 127; and a light chain variable region comprising:(d) a CDRL1 selected from the group consisting of SEQ ID NO: 128 to SEQ ID NO: 168;(e) a CDRL2 selected from the group consisting of SEQ ID NO: 169 to SEQ ID NO: 192; and(f) a CDRL3 selected from the group consisting of SEQ ID NO: 193 to SEQ ID NO: 238.

2. The antibody, or antigen-binding fragment thereof, of claim 1 , wherein the heavy chain variable region comprise CDRs selected from one of the following:(a) CDRHs according to SEQ ID NO: 2, 39 and 79;(b) CDRHs according to SEQ ID NO: 1 , 38 and 78;(c) CDRHs according to SEQ ID NO: 3, 40 and 80;(d) CDRHs according to SEQ ID NO: 9, 74 and 123;(e) CDRHs according to SEQ ID NO: 6, 43 and 83;(f) CDRHs according to SEQ ID NO: 16, 52 and 93;(g) CDRHs according to SEQ ID NO: 19, 54 and 96;(h) CDRHs according to SEQ ID NO: 8, 44 and 85;(i) CDRHs according to SEQ ID NO: 12, 48 and 89;(j) CDRHs according to SEQ ID NO: 13, 64 and 107;(k) CDRHs according to SEQ ID NO: 33, 54 and 118;(l) CDRHs according to SEQ ID NO: 13, 40 and 121 ;(m) CDRHs according to SEQ ID NO: 13, 76 and 125; or(n) CDRHs according to SEQ ID NO: 37, 40 and 126.

3. The antibody, or antigen-binding fragment thereof, of claim 1 or 2, wherein the light chain variable region comprises CDRs selected from one of the following:(a) CDRLs according to SEQ ID NO: 129, 170 and 194;(b) CDRLs according to SEQ ID NO: 128, 169 and 193;(c) CDRLs according to SEQ ID NO: 130, 171 and 195;(d) CDRLs according to SEQ ID NO: 143, 190 and 234;(e) CDRLs according to SEQ ID NO: 133, 178 and 198;(f) CDRLs according to SEQ ID NO: 142, 171 and 207;(g) CDRLs according to SEQ ID NO: 145, 182 and 210;(h) CDRLs according to SEQ ID NO: 135, 175 and 200;(i) CDRLs according to SEQ ID NO: 139, 179 and 203;(j) CDRLs according to SEQ ID NO: 154, 169 and 220;(k) CDRLs according to SEQ ID NO: 143, 181 and 229;(l) CDRLs according to SEQ ID NO: 165, 171 and 232;(m) CDRLs according to SEQ ID NO: 143, 171 and 236; or(n) CDRLs according to SEQ ID NO: 162, 191 and 237.

4. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 1 , 38 and 78 and CDRLs according to SEQ ID NO: 128, 169 and 193.

5. The antibody, or antigen-binding fragment thereof, according to claim 4, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 239, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 289.

6. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 2, 39 and 79 and CDRLs according to SEQ ID NO: 129, 170 and 194.

7. The antibody, or antigen-binding fragment thereof, according to claim 6, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 240, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 290.

8. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 3, 40 and 80 and CDRLs according to SEQ ID NO: 130, 171 and 195.

9. The antibody, or antigen-binding fragment thereof, according to claim 8, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO 241 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 291.

10. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 9, 74 and 123 and CDRLs according to SEQ ID NO: 143, 190 and 234.

11. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 6, 43 and 83 and CDRLs according to SEQ ID NO: 133, 178 and 198.

12. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 16, 52 and 93 and CDRLs according to SEQ ID NO: 142, 171 and 207.

13. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 19, 54 and 96 and CDRLs according to SEQ ID NO: 145, 182 and 210.

14. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 8, 44 and 85 and CDRLs according to SEQ ID NO: 135, 175 and 200.

15. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 12, 48 and 89 and CDRLs according to SEQ ID NO: 139, 179 and 203.

16. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 64 and 107 and CDRLs according to SEQ ID NO: 154, 169 and 220.

17. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 33, 54 and 118 and CDRLs according to SEQ ID NO: 143, 181 and 229.

18. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 40 and 121 and CDRLs according to SEQ ID NO: 165, 171 and 232.

19. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 13, 76 and 125 and CDRLs according to SEQ ID NO: 143, 171 and 236.

20. The antibody, or antigen-binding fragment thereof, according to any of claims 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 37, 40 and 126 and CDRLs according to SEQ ID NO: 162, 191 and 237.

21. The antibody, or antigen-binding fragment thereof, of any one of the preceding claims, wherein the antibody binds to HLA-G.

22. The antibody, or antigen-binding fragment thereof, of any one of the preceding claims for use in therapy.

23. The antibody, or antigen-binding fragment thereof, of any one of the preceding claims, for use in the treatment or prevention of cancer.

24. The antibody, or antigen-binding fragment thereof, of any one of the preceding claims, for use in the manufacture of a medicament for the treatment of cancer.

25. A nucleic acid encoding an antibody, or an antigen-binding fragment thereof, according to any one of the preceding claims.

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