Anti-lilrb2 antibodies and their use in the treatment of cancer

Abstract

The present invention relates to antibodies, or antigen-binding fragments thereof, that are capable of binding to Leukocyte Immunoglobulin-Like Receptor B2 (LILRB2). The present invention also relates to methods of producing and therapeutic uses of such antibodies, or antigen-binding fragments thereof.

Description

[0001] ANTIBODY

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to antibodies, or antigen-binding fragments thereof, that are capable of binding to Leukocyte Immunoglobulin-Like Receptor B2 (LILRB2). The present invention also relates to methods of producing and therapeutic uses of such antibodies, or antigen-binding fragments thereof.

[0004] BACKGROUND

[0005] Antibodies are glycoproteins belonging to the immunoglobulin superfamily of proteins. There are five primary classes of immunoglobulin, including IgG, IgM, IgA, IgE and IgD. Antibodies are typically made of basic structural units comprising two heavy and two 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] LILRB2 contains four tandem Ig-like domains and is a receptor for classical and non-classical MHC- 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 LILRB2 binding domains that bind specifically to LILRB2 and not the other LILRs (such as LILRA1 , LILRA2 and LILA5).

[0016] The present inventors have developed an antibody, or antigen-binding fragment thereof, that targets LILRB2 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: 39;

[0019] (b) a CDRH2 selected from the group consisting of SEQ ID NO 40 to SEQ ID NO 80; and

[0020] (c) a CDRH3 selected from the group consisting of SEQ ID NO 81 to SEQ ID NO 130; and a light chain variable region comprising:

[0021] (d) a CDRL1 selected from the group consisting of SEQ ID NO: 131 to SEQ ID NO: 164;

[0022] (e) a CDRL2 selected from the group consisting of SEQ ID NO: 165 to SEQ ID NO: 184; and

[0023] (f) a CDRL3 selected from the group consisting of SEQ ID NO: 185 to SEQ ID NO: 228.

[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 , 40 and 81;

[0026] (b) CDRHs according to SEQ ID NO: 2, 41 and 82;

[0027] (c) CDRHs according to SEQ ID NO: 3, 42 and 83;

[0028] (d) CDRHs according to SEQ ID NO: 7, 45 and 87;

[0029] (e) CDRHs according to SEQ ID NO: 12, 50 and 93;

[0030] (f) CDRHs according to SEQ ID NO: 4, 43 and 84;

[0031] (g) CDRHs according to SEQ ID NO: 8, 50 and 88;

[0032] (h) CDRHs according to SEQ ID NO: 7, 46 and 89; (i) CDRHs according to SEQ ID NO: 25, 65 and 113;

[0033] (j) CDRHs according to SEQ ID NO: 36, 78 and 127;

[0034] (k) CDRHs according to SEQ ID NO: 6, 44 and 86;

[0035] (l) CDRHs according to SEQ ID NO: 30, 73 and 121 ;

[0036] (m) CDRHs according to SEQ ID NO: 38, 47 and 129;

[0037] (n) CDRHs according to SEQ ID NO: 7, 72 and 120;

[0038] (o) CDRHs according to SEQ ID NO: 14, 53 and 96; or

[0039] (p) CDRHs according to SEQ ID NO: 5, 57 and 102.

[0040] 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:

[0041] (a) CDRLs according to SEQ ID NO: 131 , 165 and 185;

[0042] (b) CDRLs according to SEQ ID NO: 132, 166 and 186;

[0043] (c) CDRLs according to SEQ ID NO: 133, 165and 187;

[0044] (d) CDRLs according to SEQ ID NO: 137, 168 and 191 ;

[0045] (e) CDRLs according to SEQ ID NO: 136, 167 and 197;

[0046] (f) CDRLs according to SEQ ID NO: 134, 166 and 188;

[0047] (g) CDRLs according to SEQ ID NO: 133, 169 and 192;

[0048] (h) CDRLs according to SEQ ID NO: 138, 170 and 193; (i) CDRLs according to SEQ ID NO: 155, 179 and 203;

[0049] (j) CDRLs according to SEQ ID NO: 161 , 168 and 226;

[0050] (k) CDRLs according to SEQ ID NO: 133, 168 and 190;

[0051] (l) CDRLs according to SEQ ID NO: 164, 168 and 221 ;

[0052] (m) CDRLs according to SEQ ID NO: 163, 168 and 227;

[0053] (n) CDRLs according to SEQ ID NO: 164, 179 and 220;

[0054] (o) CDRLs according to SEQ ID NO: 142, 172 and 200; or

[0055] (p) CDRLs according to SEQ ID NO: 164, 165 and 206.

[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: 1 , 40 and

[0057] 81 and CDRLs according to SEQ ID NO: 131, 165 and 185.

[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: 229, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 279.

[0059] 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, 41 and

[0060] 82 and CDRLs according to SEQ ID NO: 132, 166 and 186.

[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: 230, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 280.

[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: 3, 42 and 83 and CDRLs according to SEQ ID NO: 133, 165 and 187. 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: 231 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 281.

[0063] 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: 7, 45 and 87 and CDRLs according to SEQ ID NO: 137, 168 and 191.

[0064] 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: 232, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 282.

[0065] 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, 50 and 93 and CDRLs according to SEQ ID NO: 136, 167 and 197.

[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: 234, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 284.

[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: 4, 43 and 84 and CDRLs according to SEQ ID NO: 134, 166 and 188.

[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: 235, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 285.

[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: 8, 50 and 88 and CDRLs according to SEQ ID NO: 133, 169 and 192. 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: 238, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 288.

[0070] 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: 7, 46 and 89 and CDRLs according to SEQ ID NO: 138, 170 and 193.

[0071] 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.

[0072] 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: 25, 65 and 113 and CDRLs according to SEQ ID NO: 155, 179 and 203.

[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: 262, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 312.

[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: 36, 78 and 127 and CDRLs according to SEQ ID NO: 161, 168 and 226.

[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: 276, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 326.

[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: 6, 44 and 86 and CDRLs according to SEQ ID NO: 133, 168 and 190. 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: 237, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 287.

[0077] 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: 30, 73 and 121 and CDRLs according to SEQ ID NO: 164, 168 and 221.

[0078] 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: 270, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 320.

[0079] 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: 38, 47 and 129 and CDRLs according to SEQ ID NO: 163, 168 and 227.

[0080] 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: 278, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 328.

[0081] 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: 7, 72 and 120 and CDRLs according to SEQ ID NO: 164, 179 and 220.

[0082] 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.

[0083] 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: 14, 53 and 96 and CDRLs according to SEQ ID NO: 142, 172 and 200. 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: 245, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 295.

[0084] 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: 5, 57, 102 and CDRLs according to SEQ ID NO: 164, 165 and 206.

[0085] 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: 251 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 301.

[0086] 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.

[0087] BRIEF DESCRIPTION OF FIGURES

[0088] Figure 1 : On-target flow cytometric analysis of mAb binding to LILRB2-expressing HEK293T cells. Cells transfected with LILRB2 were stained with anti-LILRB2 mAb clones (thick line, open histogram) or human lgG4 isotype control (grey shaded histogram), followed by anti-human lgG4- APC secondary antibody. Confirmation of LILRB2 expression by HEK293T was demonstrated using an APC-conjugated commercial anti-LILRB2 mAb.

[0089] Figure 2: Off-target flow cytometric analysis of mAb binding to LILRB-expressing HEK293T cells. Cells transfected with LILRB1, LILRB3, LILRB4 or LILRB5were stained with anti-LILRB2 mAb clones or human lgG4 isotype control, followed by anti-human IgG-APC secondary antibody. Data is presented as MFI of anti-LILRB2 stain relative to isotype control.

[0090] Figure 3: Off-target flow cytometric analysis of mAb binding to LILRA-expressing HEK293T cells. Cells transfected with LILRA1 , LILRA2, LILRA5 or LILRA6 were stained with anti-LILRB2 mAb clones or human lgG4 isotype control, followed by anti-human IgG-APC secondary antibody. Data is presented as FI of anti-LILRB2 stain relative to isotype control.

[0091] Figure 4: Direct ELISA. Data shows anti-LILRB2 mAbs bind to recombinant LILRB2-FC. The lower the EC50 the better binding the anti-LILRB2 is to recombinant LILRB2-FC.

[0092] Figure 5: Competitive ELISA. Data shows anti-LILRB2 antibodies that block the binding of recombinant HLAG to recombinant LILRB2-Fc. The higher the percent blocking the better the capacity to block binding of the two recombinant antigens.

[0093] Figure 6: Expression and Purity. Data shows that mAb clones demonstrate expression above 200 mg / kg and acceptable purity.

[0094] Figure 7: Thermostability. Data shows each mAb has good thermostability representing positive chemistry, manufacturing and controls (CMC) attributes.

[0095] Figure 8: TNF-a 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: KB16A.41.2 Binding to Human LILRB2 by ELISA. KB16A.41.2 binds to recombinant human LILRB2 with a strong affinity (0.06321 pg / mL) comparable to MK-4830 produced in Nicotiana benthamiana with an affinity of 0.06972 pg / mL.

[0096] Figure 10: KB16A.41.2 Human vs Cynomolgus LILRB2 Binding. KB16A.41.2 did not demonstrate binding affinity for Cynomolgus LILRB2, but bound with high affinity to human LILRB2. Antibody MK-4830 produced in Nicotiana benthamiana was used as a positive binding control.

[0097] Figure 11 : KB16A.41.2 Binding to Human LILRB2 vs LILR-family members. KB16A.41.2 binds to LILRB2 expressed on the surface of HEK293T cells while there is no detectable binding of KB16A.41.2 to LILRB1 , LILRB3, LILRB4, LILRB5, LILRA1, LILRA5 or LILRA6.

[0098] Figure 12: KB16A.41.2 Binding to HEK293T cells expressing LILRB2. KB16A.41.2 binds to LILRB2 over-expressed on the surface of HEK293T cells with an EC5o of 0.021 pg / mL and does not bind to HEK293T cells untransfected or transfected with plasmid expressing LILRB1.

[0099] Figure 13: KB16A.41.2 Binding to primary human monocytes. KB16A.41.2 binds to primary human CD14 monocytes with an ECso of 0.05132 pg / mL.

[0100] Figure 14: Binding Confirmation of KB16A.41.2 by Surface Plasmon Resonance (SPR). KB16A.41.2 binds to LILRB2 with high affinity and corroborates the ELISA and flow cytometry binding data.

[0101] Figure 15: LILRB2 / HLA-G Blocking ELISA. KB16A.41.2 exhibited blocking activity with an IC5o of 0.3043 pg / mL confirming that the antibody is able to block binding association between recombinant HLA-G and LILRB2. MK-4830 produced in Nicotiana benthamiana was used as a positive control and has an IC50 of 0.4172 pg / mL.

[0102] Figure 16: Prevention of HLA-G Induced Monocyte Suppression. KB16A.41.2 prevents HLA-G induced monocyte suppression of TNF-a cytokine release in a dose-dependent manner when compared to controls. Figure 8A data are expressed as mean ± SEM of two separate experiments. Suppression control had 43% less TNF-a than the stimulation control. Figure 8B is representative of one experiment and data are expressed as mean ± SD, and MK-4830 produced in Nicotiana benthamiana was used as a positive control. Suppression control had 38% less TNF-a than the stimulation control. Treatment with lgG4 isotype control results in no prevention of HLA-G mediated suppression of TNF-a with both JEG-3 and HT-1376 cell lines. Percent TNF-a release by KB16A.41.2 was calculated between treatment groups and stimulation control. The “#” indicates >80% TNF-a release was maintained in comparison to stimulation control. Figure 17: Monocyte Activation. KB16A.41.2 induces monocyte activation via increased TNF-a cytokine release in a dose-dependent manner when compared to controls. DataError! Reference source not found, are expressed as mean ± SEM of three separate experiments with one donor. Monocyte activation was replicated in two additional donors that are not shown. Treatment with KB16A.41.2 results in a significant increase in TNF-a in each treatment group and no observable change in IL10 production in this assay.

[0103] Figure 18: D6 macrophages differentiation phenotyping. KB16A.41.2 antibody significantly modulated the phenotype of differentiating macrophages, increasing pro-inflammatory marker expression (CD86) while downregulating anti-inflammatory markers (CD14, CD163, CD206, CD209). This suggests KB16A.41.2 promotes polarization towards a pro-inflammatory phenotype. Data are presented as single data points of gMFI (geometric mean fluorescence intensity) or representative histograms. Statistical analysis used paired t-tests (n=5 donors; 7 replicates; *p<0.05, **p<0.01 , ***p<0.001 ; grey: isotype control; red: KB16A.41.2).

[0104] Figure 19: D6 moDC differentiation phenotyping. KB16A.41.2 antibody modulated the phenotype of differentiating moDCs, increasing pro-inflammatory marker expression (CD86) while downregulating anti-inflammatory markers (CD14, CD163, CD206, CD209). This suggests KB16A.41.2 promotes polarization towards a pro-inflammatory phenotype. A representative histogram for each marker is shown (n=2 donors; grey: isotype control; red: KB16A.41.2).

[0105] Figure 20: T-cell-macrophage MLR. To determine their T cell activation potential, macrophages differentiated in the presence of KB16A.41.2 were used in a mixed leukocyte reaction (MLR) with naive T cells. Macrophages differentiated in the presence of KB16A.41.2 were able to elicit better T cell survival (assessed by enumerating T cells at the end of culture), increased T cell activation (assessed by activation marker CD25), and improved pro-inflammatory cytokine production (assessed by secretion of IFN-y and I L-113).

[0106] Figure 21 :Tumor volume. Mean Change (A) in SK-MEL-5 tumor volume growth. The treatment (20 mg / kg) was performed BIWfor 24 days (8 total doses). The study was terminated on Day 31. The mean (±SEM) Atumor volume from each dosing group is plotted (n=9). ATumor Volume = Vt -VO; VO is the individual’s tumor volume on the day of randomization (day 0), Vt is its tumor volume after the day of randomization (Day 1-31) on a given day (day t). Figure 22: Mean Tumor Weight. Mean whole SK-MEL-5 tumor weight at Day 31. The mean (±SEM) tumor weights (n = 9) are plotted. P<0.05 is considered statistically significant compared with “Isotype group” determined by a One-way ANOVA with a Fisher’s LSD test.

[0107] Figure 23: Absolute count(up) and Percentage(down) of cell subsets in spleen and tumor samples per flow cytometry analysis. *P < 0.05, **P < 0.01 , ***P < 0.001 is considered statistically significant compared with “Isotype group”.

[0108] Figure 24: Percentage of Human CD45+lmmune Cell Populations. Percentage of human CD45+ immune cell populations in blood, spleen, and tumor samples per flow cytometry analysis. *P < 0.05, **P < 0.01, ***P < 0.001 is considered statistically significant compared with “Isotype group”.

[0109] 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). Top left, hlgG4 Isotype Control tumor samples labeled SK-MEL-5-hHLA-G-#8-G1. Top right, KB16A.41.2 tumor samples labeled SK-MEL-5-hHLA-G-#8-G2. Bottom, Clinical Comparator samples labeled SK-MEL-5-hHLA-G-#8- G5. JEG-3 cells were used as a positive control for HLA-G expression and the parental SK-MEL-5 cell-line (no HLA-G expression) was used as a negative control for HLA-G expression.

[0110] Figure 26: Mean Tumor Volume. Mean HT-1376 tumor volume by treatment group. 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 Tukey multiple comparison test was performed to compare all groups. P values are presented as * color coordinated with the group that is being compared to Isotype control or as # color coordinated with the group that is being compared to Atezolizumab or as + color coordinated with the group that is being compared to KB16A.41.2;*P < 0.05, **P < 0.01 , ***P < 0.001..

[0111] Figure 27: Mean (A) Tumor Volume. Mean change (A) in HT-1376 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). 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. ATumor Volume = Vt -VO; VO is the individual’s tumor volume on the day of randomization (day 0), Vt is its tumor volume after the day of randomization on a given day (day t).

[0112] Figure 28: Cell Subsets in Tumor Samples. Absolute count result (top) and percentage (bottom) of cell subsets in tumor samples per flow cytometry analysis. P<0.05 is considered statistically significant compared with “Isotype group”.

[0113] DETAILED DESCRIPTION

[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 each have an amino acid sequence according to any one of SEQ ID NO: 1 to SEQ ID NO: 130, and wherein the antibody, or antigen-binding fragment thereof, comprises three complementarity determining regions on the light chain variable region (CDRLs), wherein the CDRLs each have an amino acid sequence according to any one of SEQ ID NO: 131 to SEQ ID NO: 228.

[0116] 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:

[0117] (a) a CDRH1 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 39;

[0118] (b) a CDRH2 selected from the group consisting of SEQ ID NO 40 to SEQ ID NO 80; and

[0119] (c) a CDRH3 selected from the group consisting of SEQ ID NO 81 to SEQ ID NO 130; and a light chain variable region comprising:

[0120] (d) a CDRL1 selected from the group consisting of SEQ ID NO: 131 to SEQ ID NO: 164;

[0121] (e) a CDRL2 selected from the group consisting of SEQ ID NO: 165 to SEQ ID NO: 184; and

[0122] (f) a CDRL3 selected from the group consisting of SEQ ID NO: 185 to SEQ ID NO: 228.

[0123] 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:

[0124]

[0125]

[0126]

[0127] 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:

[0128]

[0129] 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.

[0130] 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:

[0131]

[0132] 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:

[0133] The KB identifiers provided in Tables 1-4 correspond to the KB identifiers provided in the Figures and Examples. For example, KB16A.41 sequences relate to KB16A.41.2 in the figures.

[0134] 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: 229 to 278 and a light chain variable region as defined by any of SEQ ID NOs: 279 to 328.

[0135] 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,

[0136] 40 and 81 and light chain variable region comprising CDRLs according to SEQ ID NO: 131, 165 and

[0137] 185.

[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: 229, 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: 279.

[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: 2,

[0140] 41 and 82 and light chain variable region comprising CDRLs according to SEQ ID NO: 132, 166 and

[0141] 186.

[0142] 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: 230, 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: 280.

[0143] 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, 42 and 83 and light chain variable region comprising CDRLs according to SEQ ID NO: 133, 165 and 187.

[0144] 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: 231, 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: 281.

[0145] 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, 45 and 87 and light chain variable region comprising CDRLs according to SEQ ID NO: 137, 168 and 191.

[0146] 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: 232, 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: 282.

[0147] 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, 50 and 93 and light chain variable region comprising CDRLs according to SEQ ID NO: 136, 167 and 197.

[0148] 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: 234, 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: 284.

[0149] 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, 43 and 84 and light chain variable region comprising CDRLs according to SEQ ID NO: 134, 166 and 188.

[0150] 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: 235, 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: 285.

[0151] 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, 50 and 88 and light chain variable region comprising CDRLs according to SEQ ID NO: 133, 169 and

[0152] 192.

[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: 238, 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: 288.

[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: 7, 46 and 89 and light chain variable region comprising CDRLs according to SEQ ID NO: 138, 170 and

[0155] 193.

[0156] 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: 25, 65 and 113 and light chain variable region comprising CDRLs according to SEQ ID NO: 155, 179 and 203.

[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: 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.

[0158] 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, 78 and 127 and light chain variable region comprising CDRLs according to SEQ ID NO: 161 , 168 and 226.

[0159] 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.

[0160] 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, 44 and 86 and light chain variable region comprising CDRLs according to SEQ ID NO: 133, 168 and 190.

[0161] 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: 237, 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: 287.

[0162] 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, 73 and 121 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 168 and 221.

[0163] 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.

[0164] 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: 38, 47 and 129 and light chain variable region comprising CDRLs according to SEQ ID NO: 163, 168 and 227. 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.

[0165] 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, 72 and 120 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 179 and 220.

[0166] 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.

[0167] 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, 53 and 96 and light chain variable region comprising CDRLs according to SEQ ID NO: 142, 172 and 200.

[0168] 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.

[0169] 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, 57, 102 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 165 and 206.

[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: 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.

[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: 39, 80 and 130 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 165 and 228.

[0172] 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: 233, 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: 283.

[0173] 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, 50 and 85 and light chain variable region comprising CDRLs according to SEQ ID NO: 135, 165 and 189.

[0174] 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: 236, 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: 286.

[0175] 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, 47 and 90 and light chain variable region comprising CDRLs according to SEQ ID NO: 139, 168 and 194.

[0176] 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.

[0177] 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,

[0178] 48 and 91 and light chain variable region comprising CDRLs according to SEQ ID NO: 140, 165 and

[0179] 195.

[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: 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.

[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: 11 ,

[0182] 49 and 92 and light chain variable region comprising CDRLs according to SEQ ID NO: 137, 171 and

[0183] 196.

[0184] 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. 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,

[0185] 51 and 94 and light chain variable region comprising CDRLs according to SEQ ID NO: 141, 166 and

[0186] 198.

[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: 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.

[0188] 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,

[0189] 52 and 95 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 168 and

[0190] 199.

[0191] 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.

[0192] 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, 80 and 97 and light chain variable region comprising CDRLs according to SEQ ID NO: 143, 173 and 201.

[0193] 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.

[0194] 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,

[0195] 54 and 98 and light chain variable region comprising CDRLs according to SEQ ID NO: 138, 170 and

[0196] 202.

[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: 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.

[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: 17,

[0199] 55 and 99 and light chain variable region comprising CDRLs according to SEQ ID NO: 144, 174,

[0200] 203. 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.

[0201] 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, 56 and 100 and light chain variable region comprising CDRLs according to SEQ ID NO: 145, 165 and 204.

[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: 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.

[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: 19, 45 and 101 and light chain variable region comprising CDRLs according to SEQ ID NO: 146, 175 and 205.

[0204] 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.

[0205] 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, 58 and 103 and light chain variable region comprising CDRLs according to SEQ ID NO: 147, 165 and 207.

[0206] 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.

[0207] 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, 50 and 104 and light chain variable region comprising CDRLs according to SEQ ID NO: 148, 168 and 208.

[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: 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.

[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: 7, 50 and 105 and light chain variable region comprising CDRLs according to SEQ ID NO: 149, 168 and 209.

[0210] 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.

[0211] 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, 50 and 106 and light chain variable region comprising CDRLs according to SEQ ID NO: 135, 173 and 210.

[0212] 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.

[0213] 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,

[0214] 59 and 107 and light chain variable region comprising CDRLs according to SEQ ID NO: 150, 176 and 211.

[0215] 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.

[0216] 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,

[0217] 60 and 108 and light chain variable region comprising CDRLs according to SEQ ID NO: 151 , 174 and 203.

[0218] 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. 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 ,

[0219] 61 and 109 and light chain variable region comprising CDRLs according to SEQ ID NO: 152, 168 and 212.

[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: 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.

[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: 22,

[0222] 62 and 110 and light chain variable region comprising CDRLs according to SEQ ID NO: 133, 177 and 213.

[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: 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.

[0224] 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, 63 and 111 and light chain variable region comprising CDRLs according to SEQ ID NO: 153, 178 and 214.

[0225] 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.

[0226] 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, 64 and 112 and light chain variable region comprising CDRLs according to SEQ ID NO: 154, 169 and 215.

[0227] 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.

[0228] 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, 66 and 114 and light chain variable region comprising CDRLs according to SEQ ID NO: 133, 168 and 216. 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.

[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: 27,

[0230] 67 and 115 and light chain variable region comprising CDRLs according to SEQ ID NO: 138, 170 and 217.

[0231] 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.

[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: 11 ,

[0233] 68 and 116 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 168 and 216.

[0234] 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.

[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: 28,

[0236] 69 and 117 and light chain variable region comprising CDRLs according to SEQ ID NO: 156, 180 and 218.

[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: 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.

[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: 29,

[0239] 70 and 118 and light chain variable region comprising CDRLs according to SEQ ID NO: 157, 181 and 219.

[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: 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.

[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: 7, 71 and 119 and light chain variable region comprising CDRLs according to SEQ ID NO: 158, 168 and 195.

[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: 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.

[0243] 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 , 74 and 122 and light chain variable region comprising CDRLs according to SEQ ID NO: 164, 182 and 195.

[0244] 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.

[0245] 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, 50 and 123 and light chain variable region comprising CDRLs according to SEQ ID NO: 159, 168 and 222.

[0246] 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.

[0247] 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, 75 and 124 and light chain variable region comprising CDRLs according to SEQ ID NO: 160, 183 and 223.

[0248] 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: 34,

[0249] 76 and 125 and light chain variable region comprising CDRLs according to SEQ ID NO: 135, 182 and 224.

[0250] 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.

[0251] 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,

[0252] 77 and 126 and light chain variable region comprising CDRLs according to SEQ ID NO: 158, 184 and 225.

[0253] 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.

[0254] 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, 79 and 128 and light chain variable region comprising CDRLs according to SEQ ID NO: 162, 174 and 225.

[0255] 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.

[0256] 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:

[0257] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSEAMSWVRQAPGKGLEWVSAIGAGGGTYYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARIVSGWTGDAFDIWGQGTMVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 329)

[0258] 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: DIQMTQSPSSLSASVGDRVTITCRASQSIGSSLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCQQSYSTPQYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 330)

[0259] 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: 329, 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: 330.

[0260] Antibody Binding Properties

[0261] The antibody of the invention, or antigen-binding fragment thereof, may be defined by its binding specificity.

[0262] In one embodiment, the antibody, or antigen-binding fragment thereof, binds specifically to LILRB2. In other words, the antibody, or antigen-binding fragment thereof, does not bind or bind to a lesser extent to other LILRs.

[0263] Binding specificity may be measured through a LILRB2 On-Target Screen and a LILRB2 Off-Target Screen as detailed in the Examples.

[0264] 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.

[0265] Direct ELISA is used herein to measure the EC50 of anti-LILRB2 mAbs to recombinant human LILRB2 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.

[0266] 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.

[0267] 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.

[0268] 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.

[0269] 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.

[0270] Antibody binding may also be measured through a competitive ELISA. This assay is used herein to measure how much the anti-LILRB2 antibodies disclosed herein may block the binding of recombinant HLA-G to recombinant LILRB2.

[0271] 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.

[0272] 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.

[0273] 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.

[0274] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 20% blocking of HLA-G and LILRB2 interaction. Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 30% blocking of HLA-G and LILRB2 interaction.

[0275] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 40% blocking of HLA-G and LILRB2 interaction.

[0276] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 50% blocking of HLA-G and LILRB2 interaction.

[0277] Suitably, the antibody, or antigen-binding fragment thereof, as disclosed herein may achieve at least about 60% blocking of HLA-G and LILRB2 interaction.

[0278] Antibody Thermostability

[0279] 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.

[0280] In one embodiment, the antibody, or antigen-binding fragment thereof, is thermostable up 60°C.

[0281] In one embodiment, the antibody, or antigen-binding fragment thereof, is thermostable up 70°C.

[0282] In one embodiment, the antibody, or antigen-binding fragment thereof, is thermostable up 80°C.

[0283] Additional features of antibody or antigen-binding fragment thereof

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

[0285] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG1 and all allotypes thereof. In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG2 and all allotypes thereof.

[0286] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG3 and all allotypes thereof.

[0287] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgG4 and all allotypes thereof.

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

[0289] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgA1 and allotypes thereof.

[0290] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgA2 and all allotypes thereof.

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

[0292] In one embodiment the antibody, or antigen-binding fragment thereof, may be IgE and all allotypes thereof.

[0293] In one embodiment the antibody, or antigen-binding fragment thereof, may be IgD and all isotypes thereof.

[0294] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgD1 and all allotypes thereof.

[0295] In one embodiment the antibody, or antigen-binding fragment thereof, may be lgD2 and all allotypes thereof.

[0296] In one embodiment, the antibody, or antigen-binding fragment thereof is a chimeric antibody.

[0297] In one embodiment, the antibody, or antigen-binding fragment thereof, is a humanized antibody.

[0298] In one embodiment, the antibody, or antigen-binding fragment thereof is an affinity-matured antibody.

[0299] In one embodiment, the antibody, or antigen-binding fragment thereof, is an affinity-matured, humanized antibody.

[0300] 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 I gG 1 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.

[0301] Antibody fragments

[0302] 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).

[0303] 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.

[0304] 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.

[0305] 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.

[0306] In one embodiment, the antigen-binding fragment of the antibodies disclosed herein may be any fragment of the antibodies disclosed herein.

[0307] 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.

[0308] 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. 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).

[0309] Pharmaceutical formulations

[0310] 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.

[0311] 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.

[0312] 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.

[0313] Medical use

[0314] 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.

[0315] 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.

[0316] 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 the manufacture of a medicament.

[0317] 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. In one embodiment, the present invention provides an antibody, or antigen-binding fragment thereof, as disclosed herein for use as an anti-cancer therapeutic.

[0318] 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.

[0319] 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.

[0320] 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.

[0321] 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.

[0322] 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.

[0323] 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.

[0324] 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.

[0325] 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.

[0326] 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. 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.

[0327] 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.

[0328] 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.

[0329] 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.

[0330] LILRB2 may be expressed on immune cells, such as myeloid cells.

[0331] LILRB2 may be expressed on immune cells, such as monocytes, macrophages, granulocytes.

[0332] LILRB2 may be expressed on immune cells such as, T-cells.

[0333] In some embodiments, the subject of the medical uses and methods of treatment according to the present invention may be a mammal.

[0334] In some embodiments, the subject may be a human.

[0335] In some embodiments, the subject may be a patient, such as a human patient.

[0336] In some embodiments, the subject may suffer from and / or have been diagnosed with cancer.

[0337] 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. In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of cancer.

[0338] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of solid tumours.

[0339] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of bladder cancer.

[0340] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of placental cancer.

[0341] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment or prevention of melanoma.

[0342] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment and prevention of cancer, in combination with an additional anti-cancer therapeutic.

[0343] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment and prevention of cancer, in combination with an additional anti-cancer antibody.

[0344] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment and prevention of cancer, in combination with an anti-PD-L1 antibody.

[0345] In some embodiments, the antibody, or antigen-binding fragment thereof, as disclosed herein may be used in the treatment and prevention of cancer, in combination with Atezolizumab.

[0346] Antibody-drug conjugates

[0347] 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.

[0348] In one embodiment, the present invention provides an antibody-drug conjugate comprising the antibody, or antigen-binding fragment thereof, disclosed herein.

[0349] 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.

[0350] 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.

[0351] 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.

[0352] 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.

[0353] 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.

[0354] 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.

[0355] 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.

[0356] 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. Antibody-Toxin Fusion Proteins

[0357] An antibody-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 antibody-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.

[0358] 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.

[0359] 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.

[0360] 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.

[0361] 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.

[0362] Nucleotide Sequences

[0363] The antibody, or antigen-binding fragment thereof, may be encoded by any suitable nucleotide sequence.

[0364] 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.

[0365] 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.

[0366] In one embodiment, the present invention provides a nucleic acid that encodes an antibody, or antigen-binding fragment thereof, as disclosed herein.

[0367] 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.

[0368] Expression System

[0369] The present invention further relates to a method of production and engineering of antibodies, or antigen-binding fragments thereof, disclosed herein.

[0370] 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.

[0371] 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.

[0372] 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. 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.

[0373] 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.

[0374] Further Embodiments

[0375] 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.

[0376] 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.

[0377] 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. 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.

[0378] Numbered paragraphs

[0379] 1. An antibody, or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising:

[0380] (a) a CDRH1 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 39;

[0381] (b) a CDRH2 selected from the group consisting of SEQ ID NO 40 to SEQ ID NO 80; and

[0382] (c) a CDRH3 selected from the group consisting of SEQ ID NO 81 to SEQ ID NO 130; and a light chain variable region comprising:

[0383] (d) a CDRL1 selected from the group consisting of SEQ ID NO: 131 to SEQ ID NO: 164;

[0384] (e) a CDRL2 selected from the group consisting of SEQ ID NO: 165 to SEQ ID NO: 184; and

[0385] (f) a CDRL3 selected from the group consisting of SEQ ID NO: 185 to SEQ ID NO: 228.

[0386] 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:

[0387] (a) CDRHs according to SEQ ID NO: 1 , 40 and 81;

[0388] (b) CDRHs according to SEQ ID NO: 2, 41 and 82;

[0389] (c) CDRHs according to SEQ ID NO: 3, 42 and 83;

[0390] (d) CDRHs according to SEQ ID NO: 7, 45 and 87;

[0391] (e) CDRHs according to SEQ ID NO: 12, 50 and 93;

[0392] (f) CDRHs according to SEQ ID NO: 4, 43 and 84;

[0393] (g) CDRHs according to SEQ ID NO: 8, 50 and 88;

[0394] (h) CDRHs according to SEQ ID NO: 7, 46 and 89;

[0395] (i) CDRHs according to SEQ ID NO: 25, 65 and 113;

[0396] (j) CDRHs according to SEQ ID NO: 36, 78 and 127;

[0397] (k) CDRHs according to SEQ ID NO: 6, 44 and 86;

[0398] (l) CDRHs according to SEQ ID NO: 30, 73 and 121 ;

[0399] (m) CDRHs according to SEQ ID NO: 38, 47 and 129;

[0400] (n) CDRHs according to SEQ ID NO: 7, 72 and 120;

[0401] (o) CDRHs according to SEQ ID NO: 14, 53 and 96; or

[0402] (p) CDRHs according to SEQ ID NO: 5, 57 and 102.

[0403] 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: (a) CDRLs according to SEQ ID NO: 131 , 165 and 185;

[0404] (b) CDRLs according to SEQ ID NO: 132, 166 and 186;

[0405] (c) CDRLs according to SEQ ID NO: 133, 165 and 187;

[0406] (d) CDRLs according to SEQ ID NO: 137, 168 and 191 ;

[0407] (e) CDRLs according to SEQ ID NO: 136, 167 and 197;

[0408] (f) CDRLs according to SEQ ID NO: 134, 166 and 188;

[0409] (g) CDRLs according to SEQ ID NO: 133, 169 and 192;

[0410] (h) CDRLs according to SEQ ID NO: 138, 170 and 193;

[0411] (i) CDRLs according to SEQ ID NO: 155, 179 and 203;

[0412] (j) CDRLs according to SEQ ID NO: 161 , 168 and 226;

[0413] (k) CDRLs according to SEQ ID NO: 133, 168 and 190;

[0414] (l) CDRLs according to SEQ ID NO: 164, 168 and 221 ;

[0415] (m) CDRLs according to SEQ ID NO: 163, 168 and 227;

[0416] (n) CDRLs according to SEQ ID NO: 164, 179 and 220;

[0417] (o) CDRLs according to SEQ ID NO: 142, 172 and 200; or

[0418] (p) CDRLs according to SEQ ID NO: 164, 165 and 206.

[0419] 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 , 40 and 81 and CDRLs according to SEQ ID NO: 131 , 165 and 185.

[0420] 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: 229, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 279.

[0421] 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, 41 and 82 and CDRLs according to SEQ ID NO: 132, 166 and 186.

[0422] 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: 230, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 280.

[0423] 8. 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, 42 and 83 and CDRLs according to SEQ ID NO: 133, 165 and 187. 9. The antibody, or antigen-binding fragment thereof, according to paragraph 8, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 231 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 281.

[0424] 10. 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: 7, 45 and 87 and CDRLs according to SEQ ID NO: 137, 168 and 191.

[0425] 11. The antibody, or antigen-binding fragment thereof, according to paragraph 10, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 232, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 282.

[0426] 12. 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, 50 and 93 and CDRLs according to SEQ ID NO: 136, 167 and 197.

[0427] 13. The antibody, or antigen-binding fragment thereof, according to paragraph 12, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 234, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 284.

[0428] 14. 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: 4, 43 and 84 and CDRLs according to SEQ ID NO: 134, 166 and 188.

[0429] 15. The antibody, or antigen-binding fragment thereof, according to paragraph 14, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 235, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 285.

[0430] 16. 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, 50 and 88 and CDRLs according to SEQ ID NO: 133, 169 and 192. 17. The antibody, or antigen-binding fragment thereof, according to paragraph 16, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 238, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 288.

[0431] 18. The antibody, or antigen-binding fragment thereof, according to any of paragraph 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 7, 46 and 89 and CDRLs according to SEQ ID NO: 138, 170 and 193.

[0432] 19. The antibody, or antigen-binding fragment thereof, according to paragraph 18, 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.

[0433] 20. 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: 25, 65 and 113 and CDRLs according to SEQ ID NO: 155, 179 and 203.

[0434] 21. The antibody, or antigen-binding fragment thereof, according to paragraph 20, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 262, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 312.

[0435] 22. The antibody, or antigen-binding fragment thereof, according to any of paragraph 1 to 3, wherein the variable region comprises CDRHs according to SEQ ID NO: 36, 78 and 127 and CDRLs according to SEQ ID NO: 161 , 168 and 226.

[0436] 23. The antibody, or antigen-binding fragment thereof, according to paragraph 22, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 276, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 326.

[0437] 24. 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, 44 and 86 and CDRLs according to SEQ ID NO: 133, 168 and 190. 25. The antibody, or antigen-binding fragment thereof, according to paragraph 24, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 237, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 287.

[0438] 26. 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: 30, 73 and 121 and CDRLs according to SEQ ID NO: 164, 168 and 221.

[0439] 27. The antibody, or antigen-binding fragment thereof, according to paragraph 26, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 270, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 320.

[0440] 28. 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: 38, 47 and 129 and CDRLs according to SEQ ID NO: 163, 168 and 227.

[0441] 29. The antibody, or antigen-binding fragment thereof, according to paragraph 28, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 278, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 328.

[0442] 30. 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: 7, 72 and 120 and CDRLs according to SEQ ID NO: 164, 179 and 220.

[0443] 31. The antibody, or antigen-binding fragment thereof, according to paragraph 30, 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.

[0444] 32. 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: 14, 53 and 96 and CDRLs according to SEQ ID NO: 142, 172 and 200. 33. The antibody, or antigen-binding fragment thereof, according to paragraph 32, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 245, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 295.

[0445] 34. 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: 5, 57, 102 and CDRLs according to SEQ ID NO: 164, 165 and 206.

[0446] 35. The antibody, or antigen-binding fragment thereof, according to paragraph 34, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 251 , and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 301.

[0447] 36. The antibody, or antigen-binding fragment thereof, of any one of paragraphs 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 , 33 or 35 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.

[0448] 37. 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).

[0449] 38. 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.

[0450] 39. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, wherein the antibody binds to LILRB2.

[0451] 40. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs for use in therapy.

[0452] 41. The antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs, for use in the treatment or prevention of cancer.

[0453] 42. 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. 43. A method of treating cancer comprising administering the antibody, or antigen-binding fragment thereof, of any one of the preceding paragraphs.

[0454] 44. A nucleic acid encoding an antibody, or an antigen-binding fragment thereof, according to any one of the preceding paragraphs.

[0455] 45. 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.

[0456] 46. 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.

[0457] 47. 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.

[0458] 48. 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.

[0459] 49. 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.

[0460] EXAMPLES

[0461] 1. LILRB2 On-Target Screen

[0462] HEK293T cells were transfected with plasmid encoding LILRB2 using the Lipofectamine 3000. Two days after transfection, cells were harvested and stained sequentially with viability dye, primary antibody (mAb clone or human lgG4 isotype control) at 0.45 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 fixed using True-Nuclear fixation reagents (Biolegend) before acquisition on NovoCyte 3005 flow cytometer. Data was analyzed using FlowJo.

[0463] 2. LILRB2 Off-Target Screen

[0464] HEK293T cells were transfected with plasmid encoding LILR-A1, -A2, -A5, -A6, -B1 , -B3, -B4, -B5 using the Lipofectamine 3000. Two days after transfection, cells were harvested and stained sequentially with viability dye, primary antibody (mAb clone or human lgG4 isotype control) at 0.45 ug / ml, and then finally with anti-human IgG-APC at 5 ug / ml. A commercial anti-HI_A-G-APC antibody was used as a positive control to demonstrate the appropriate LILR expression on the HEK293T cell line after transfection. Cell suspensions were fixed using TrueNuclear fixation reagents (Biolegend) before acquisition on NovoCyte 3005 flow cytometer. Data was analyzed using FlowJo.

[0465] 3. Direct ELISA

[0466] ELISA plates were coated with 1 pg / mL LILRB2-Fc in PBS overnight at 5 °C. 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 LILRB2-Fc- bound antibodies were detected by HRP-conjugated mouse anti human lgG4 at a 1 :30,000 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.

[0467] 4. Competitive ELISA

[0468] 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. LILRB2 mAbs at 10 pg / mL were applied onto the LILRB2-Fc coated plates and incubated for 1 h at room temperature. The biotinylated human HLAG complex tetramer (HLAG-BT) was added for each at 10, 3.33, and 1.11 pg / mL and incubated for 1 h 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.

[0469] 5. Expression and Purity 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).

[0470] 6. Thermostability

[0471] Each sample 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.

[0472] 7. JEG-3 - U937: Immunosuppression Assay

[0473] 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 the 1x EMEM media was removed and 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. LILRB2 mAbs were diluted to 2x the desired concentration in complete 1x RPMI media. 100 L of LILRB2 mAbs were added at a final concentration of 100, 1 , and 0.01 pg / mL to the JEG- 3 and U937 96-well plate. 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.

[0474] 8. Further in vitro and in vivo studies of KB16A

[0475] To support the preclinical development of KB16A, several in vitro and in vivo evaluations which are summarized below and described in more detail in the following sections. In vitro experiments were first performed to confirm binding of KB16A.41.2 with human LILRB2 and define the epitope specificity and screen for cross-reactivity between KB16A.41.2 and human LILR-family and nonhuman LILRB2. KB16A.41.2 was found to block the interaction between LILRB2 and HLAG, and cell based assays demonstrated that KB16A.41.2 prevents HLA-G mediated suppression of monocytes and antagonizes the LILRB2 receptor leading to immune agonism. Additionally, cell culture experiments demonstrate that treatment of human myeloid cells with KB16A promotes polarization to pro-inflammatory phenotypes and inflammatory cytokine production following exposure to an inflammatory stimulus.

[0476] In vivo efficacy of KB16A.41.2 has been demonstrated in two humanized (CD34+ HuHSC-NOG- EXL) murine xenograft studies. In the two studies, HLA-G expressing melanoma or bladder carcinoma cells were injected subcutaneously into mice which was followed by KB16A.41.2 treatment. The studies show that KB16A.41.2 treatment significantly reduces tumor growth and induces mechanism-driven immune cell activation in tumors of humanized mice.

[0477] The completed and planned nonclinical studies of KB16A.41.2 included herein are summarized in Table 5 below.

[0478] Table 5 Nonclinical Studies of KB16A.41.2

[0479] 8.1 In Vitro Studies of KB16 A

[0480] A panel of in vitro methods were used to define the binding affinity and specificity of the KB16A.41 .2 complementarity-determining region (CDR) for human LILRB2. These methods included ELISAs using human LILRB2 (Kactus with accession number Q8N423-1), Cynomolgus LILRB2 (Aero Biosystems, LI2-C82E3), as well as flow cytometry and surface plasmon resonance (SPR) experiments focusing on binding activity of the KB16A.41.2 CDR. These experiments demonstrated that the KB16A.41.2 CDR binds strongly and specifically to human LILRB2 expressed on the surface of human immune cells, with no cross-reactivity to human LIL-receptor family members and no observable cross-reactivity for non-human primate LILRB2. Additionally, a competitive ELISA was used to confirm that KB16A.41.2 blocks the binding of recombinant HLA-G to recombinant LILRB2- Fc.

[0481] Mechanistic evaluations of KB16A.41.2 included several in vitro and cell-based assays designed to measure the extent to which KB16A.41.2-mediated LILRB2 antagonism prevents 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). These experiments demonstrated that KB16A.41.2 treatment prevents HLA-G-induced suppression and restores immune reactivity to inflammatory stimuli in monocytes co-cultured with HLA-G-expressing cells, and increases baseline monocyte activation in the absence of HU\-G-expressing cells. Further experiments demonstrated that KB16A.41.2 treatment of differentiating leukocytes promotes polarization towards a pro-inflammatory phenotype, and that leukocytes differentiated in the presence of KB16A.41 .2 elicit a more robust T cell survival and activation when mixed with a naive T cell population.

[0482] A sequence-identical version of the MK-4830 anti-LILRB2 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.

[0483] 8.1.1 Human LILRB2 Binding

[0484] To confirm KB16A.41.2 binding to human LILRB2, ELISA plates were coated with 1 pg / mL human LILRB2-FC (Kactus, LIL-HM2B2) 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 KB16A.41 .2 starting at 10 pg / mL were applied onto the plates and incubated for 1 hour at room temperature. The human LILRB2-Fc-bound antibody was detected by HRP-conjugated mouse anti human-lgG4 at a 1:10,000 dilution. The tetramethylbenzidine 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. Antibody MK-4830 produced in Nicotiana benthamiana was used as a positive binding control. Results are shown in Figure 9 demonstrate that KB16A.41.2 binds to human LILRB2 with an ECso of 0.06321 pg / mL. 8.1.2 Non-Human Primate LILRB2 Binding

[0485] To assess KB16A.41.2 cross-reactivity to non-human LILRB2 orthologs in support of nonclinical safety species selection, a literature review of LILRB2 homology was performed. LILRB1-5 expression in myeloid cell populations appears to be primate-specific, and reagents were available to perform an ELISA-based analysis of KB16A.41.2 cross reactivity for endogenous LILRB2 in Cynomolgus macaques.

[0486] ELISA plates were coated with 1 pg / mL of either KB16A.41.2, plant-made MK-4830 positive control, or lgG4 isotype control (Leinco, LT9020) in PBS overnight at 5°C, then blocked with 5% BSA in PBST for 4 hours at room temperature. 4-fold serial dilutions starting at 1 pg / mL were applied onto the plates of biotinylated Cynomolgus (Aero Biosystems, LI2-C82E3) or Human (Kactus, HM4B2B) LILRB2 protein and incubated for 1 hour at room temperature. Bound protein was detected by HRP- conjugated streptavidin at a 1 :5,000 dilution. The tetramethylbenzidine substrate was used for detection; absorbance at 450 nm was measured on a plate reader. Graphs were plotted and EC5o’s (pg / mL) were calculated with GraphPad Prism 8.0 software. Human and Cynomolgus LILRB2 recombinant antigens have confirmed binding to the corresponding species HLAG per the vendor specification sheet.

[0487] Results are shown in Figure 10 and show that KB16A.41.2 did not demonstrate binding affinity for Cynomolgus LILRB2, but bound with high affinity to human LILRB2. Antibody MK-4830 produced in Nicotiana benthamiana was used as a positive binding control.

[0488] 8.1.3 Binding Confirmation by Flow Cytometry

[0489] Flow cytometry was used to confirm KB16A.41.2 binding to cell surface human LILRB2 and to measure cross- reactivity to LILR-family members. HEK293T cells were transfected with plasmids expressing LILRB2 (or LILR-family). After two days to allow for gene expression, cells were stained with KB16A (red histogram), isotype control (grey filled histogram), or a commercial antibody against the transfected gene (black dotted histogram) (Figure 11)Error! Reference source not found.. On- and off-target staining was performed using 0.5 pg / mL of KB16A.41.2 or isotype control. Geometric mean fluorescence intensity (gMFI) of KB16A.41.2 was compared to the isotype control.

[0490] Results are shown in Figure 11 and demonstrateError! Reference source not found, that KB16A.41.2 binds to LILRB2 expressed on the surface of HEK293T cells while there is no detectable binding of KB16A.41.2 to LILRB1, LILRB3, LILRB4, LILRB5, LILRA1 , LILRA5 or LILRA6.

[0491] Flow cytometry was then used to determine KB16A.41.2 binding affinity to cell surface human LILRB2. Transfected HEK293T cells (or untransfected cells) were harvested after two days in culture. They were stained first with KB16A.41.2 at various concentrations, followed by anti-human IgG APC-conjugated secondary antibody, and then acquired on Novocyte flow cytometer. Geometric mean fluorescence intensity (gM Fl) was plotted vs log concentration of mAb in GraphPad Prism 8.0 software to determine EC50 values.

[0492] Further results are shown in Figure 12 which demonstrates that KB16A.41.2 binds to LILRB2 overexpressed on the surface of HEK293T cells with an EC50 of 0.021 pg / mL and does not bind to HEK293T cells untransfected or transfected with plasmid expressing LILRB1.

[0493] Flow cytometry was used to confirm KB16A.41.2 binding to cell surface LILRB2 on primary human monocytes. Primary human CD14 monocytes (Stem Cell Technologies) were treated with either KB16A.41.2, MK-4830 produced in Nicotiana benthamiana as a positive control, or lgG4 isotype control (Leinco Technologies, LT9020) for 30 minutes on ice. The cells were then stained with a FITC-conjugated anti-human lgG4 secondary Ab for 1 hour on ice and washed with cold 2% FBS PBS for analysis. Cells were stained with KB16A.41 .2, isotype control (grey filled histogram) against LILRB2. Mean fluorescence intensity (MFI) of KB16A.41.2 and MK-4830 produced in Nicotiana benthamiana was compared to isotype control. Graphs were plotted and ECso’s were calculated with GraphPad Prism 8.0 Software.

[0494] Results are shown in Figure 13 and show that KB16A.41.2 binds to primary human CD14 monocytes with an EC50 of 0.05132 pg / mL.

[0495] 8. 1.4 Binding Confirmation by Surface Plasmon Resonance

[0496] The binding affinity (KD) of KB16A.41.2 to biotinylated LILRB2 (BT-LILRB2) was measured on a Biacore T200 instrument at ambient temperature. BT-LILRB2 was captured on an SA sensor chip following the manufacturer’s instructions to a surface density of about 200 RUs. Serial dilutions of KB16A.41.2 or positive control 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 positive control was MK-4830 produced in Nicotiana benthamiana. SPR was performed in triplicate and error bars represent standard deviation.

[0497] Results re shown in Figure 14 and Table 6 below, which demonstrate KB16A.41.2 binds to LILRB2 with high affinity and corroborates the ELISA and flow cytometry binding data.

[0498] Table 6 Binding Confirmation of KB16A.41.2 by SPR

[0499] 8. 1.5 KB16A.41.2 LILRB2 Blocking by ELISA

[0500] Additional ELISA experiments were performed to confirm that KB16A.41.2 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 5% BSA in PBST for 2-3 hours at room temperature. During the blocking step, a KB16A.41.2 dose curve starting at 100 pg / mL (3-fold dilutions) was prepared with 1 pg / mL HLA-G complex tetramer (HLA-G-BT) and added to the ELISA plate and incubated for one hour at room temperature. HRP-conjugated Streptavidin at a 1 :2,500 dilution was added and incubated for one hour at room temperature. The tetramethyl benzidine substrate was used for detection; absorbance at 450 nm was measured on a plate reader and IC50 was calculated in GraphPad Prism.

[0501] Results are shown in Figure 15, KB16A.41.2 exhibited blocking activity with an IC50 of 0.3043 pg / mL confirming that the antibody is able to block binding association between recombinant HLA-G and LILRB2. MK-4830 produced in Nicotiana benthamiana was used as a positive control and has an IC50 of 0.4172 pg / mL.

[0502] 8. 1.6 Prevention of HLA-G Induced Monocyte suppression

[0503] To measure KB16A.41.2’s ability to prevent HLA-G induced monocyte suppression, human monocytes were co-cultured with KB16A.41.2 in a cell-based assay as follows. Human JEG-3 cells or HT-1376 cells (ATCC), both of which endogenously express HLA-G, were plated in 96-well tissue treated plates at a density of 1.5e4 cells / well (JEG-3) or 3.0e4 cells / well (HT-1376) in complete EMEM until 90-100% confluent or 48 hours at 37°C. Additionally, cryopreserved CD14 single donor human monocyte cells (Stem Cell Technologies) were cultured in mononuclear cell media (PromoCell) for 18-24 hours prior to the assay. On the day of the assay, KB16A.41.2 and human lgG4 isotype control (Leinco Technologies, LT9020) were prepared in complete RPMI media at a 2x higher concentration for a final concentration of 400, 133, or 44 pg / mL for JEG-3 and 133 or 44 pg / mL for HT-1376 when mixed with monocytes. The EMEM was removed from JEG-3 or HT-1376 cell monolayer and CD14 monocytes were transferred to complete RPMI media. Monocytes at a density of 7.5e4 cells / well were added at the same time as the mAb mixture to the JEG-3 cells or HT-1376 cells. The plate was incubated for 20 hours at 37°C. The next day LPS-EB (Invivogen) was added at a final concentration of 100 ng / mL in RPMI media to desired wells for six hours at 37°C. Suppression (HLA-G expressing cells, monocytes, and LPS) and stimulation (monocytes and LPS) controls were included in the assay. After six 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. Graphs were plotted and all statistical analysis was performed using GraphPad Prism 8.0 software. One-way analysis of variance was used to compare the suppression control with treatment groups: *p<0.05, **p<0.01 , ***p<0.001 , ****p<0.0001.

[0504] KB16A.41.2 prevents HLA-G induced monocyte suppression of TNF-a cytokine release in a dosedependent manner when compared to controls. Figure 16A data are expressed as mean ± SEM of two separate experiments. Suppression control had 43% less TNF-a than the stimulation control. Figure 16B is representative of one experiment and data are expressed as mean ± SD, and MK- 4830 produced in Nicotiana benthamiana was used as a positive control. Suppression control had 38% less TNF-a than the stimulation control. Treatment with lgG4 isotype control results in no prevention of HLA-G mediated suppression of TNF-a with both JEG-3 and HT-1376 cell lines. Percent TNF-a release by KB16A.41.2 was calculated between treatment groups and stimulation control. The “#” indicates >80% TNF-a release was maintained in comparison to stimulation control.

[0505] 8. 1. Monocyte Activation by Cell Culture

[0506] KB16A.41.2’s ability to activate monocytes was investigated in a cell-based assay where human primary monocytes were cultured with KB16A.41.2 and stimulated with LPS as follows. Cryopreserved CD14 single donor human monocyte cells (Stem Cell Technologies) were cultured in mononuclear cell media (PromoCell) for 18-24 hours prior to the assay. After 18-24 hours KB16A.41.2 or human lgG4 isotype control (Leinco Technologies, LT9020) were prepared in complete RPMI media and added to a 96-well plate for a final concentration of 400, 133, or 44 pg / mL when mixed with monocytes. CD14 monocytes were transferred to complete RPMI media and added to the 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 added at a final concentration of 100 ng / mL in RPMI media to desired wells for six hours at 37°C. After six hours, cell culture supernatants were retained for TNF-a and IL10 ELISAs performed using TNF-a Human Uncoated ELISA kit (Thermo Fisher) and the IL10 Human Uncoated ELISA kit (Thermo Fisher) in accordance with the manufacturer’s instructions. One-way analysis of variance was used to compare the stimulation control with treatment groups: *p<0.05, ***p<0.001, ****p<0.0001. Graphs were plotted and all statistical analysis was performed using GraphPad Prism 8.0 software. KB16A.41.2 induces monocyte activation via increased TNF-a cytokine release in a dose-dependent manner when compared to controls. Figure 17 data are expressed as mean ± SEM of three separate experiments with one donor. Monocyte activation was replicated in two additional donors that are not shown. Treatment with KB16A.41.2 results in a significant increase in TNF-a in each treatment group and no observable change in IL10 production in this assay.

[0507] 8.1.8 Myeloid Differentiation and Polarization

[0508] Primary human monocytes were isolated and subsequently differentiated in vitro into either macrophages or monocyte-derived dendritic cells (moDCs). Macrophage differentiation was induced by supplementation with 40 ng / mL of macrophage colony-stimulating factor (M-CSF), while moDC differentiation was achieved through the addition of 40 ng / mL of granulocyte-macrophage colony-stimulating factor (GM-CSF) and 40 ng / ml of interleukin-4 (IL-4). The differentiation process was conducted over a 6-day period. Throughout the entire duration of the assay, cells were exposed to 1 pg / mL of either plant-expressed isotype control (lgG4) or the experimental antibody KB16A.41.2. Upon completion of the differentiation period, cells were harvested and subjected to immunophenotyping via multiparametric flow cytometry.

[0509] As shown in Figure 18, the KB16A.41.2 antibody significantly modulated the phenotype of differentiating macrophages, increasing pro-inflammatory marker expression (CD86) while downregulating anti-inflammatory markers (CD14, CD163, CD206, CD209). This suggests KB16A.41.2 promotes polarization towards a pro-inflammatory phenotype. Data are presented as single data points of gMFI (geometric mean fluorescence intensity) or representative histograms. Statistical analysis used paired t-tests (n=5 donors; 7 replicates; *p<0.05, **p<0.01 , ***p<0.001; grey: isotype control; red: KB16A.41.2).

[0510] As shown in Figure 19, the KB16A.41.2 antibody modulated the phenotype of differentiating moDCs, increasing pro-inflammatory marker expression (CD86) while downregulating anti-inflammatory markers (CD14, CD163, CD206, CD209). This suggests KB16A.41.2 promotes polarization towards a pro-inflammatory phenotype. A representative histogram for each marker is shown (n=2 donors; grey: isotype control; red: KB16A.41.2).

[0511] 8. 1.9 T-Cell-Macrophage Mixed Leukocyte Reaction (MLR)

[0512] Primary human monocytes were differentiated into macrophages over six days with 40 ng / ml M- CSF. Either KB16A.41.2 or isotype control was added to the differentiation cultures at 1 pg / ml. After six days of differentiation, macrophages were harvested and co-cultured for three days with naive T cells in the presence of 10 pg / ml anti-CD3 (clone HIT3a). Cells in the wells were counted (Moxi Orflo) and the absolute number of T cells (Figure 20A) was enumerated using the proportion of CD4+ and CD8+ cells in the well as determined by flow cytometry. Data are representative of 2 different monocyte donors. T cells co-cultured with macrophages differentiated in the presence of KB16A.41.2 or isotype control were stained for the activation marker CD25 (Figure 20B). CD25 expression on total T cells was quantified either by geometric mean fluorescence intensity (gMFI) or the percent of T cells expressing CD25. Data are representative of three monocyte donors (Figure 20B). Supernatants were harvested on day three and IFN-y and IL-13 were quantified by flow cytometry using a bead-based immunoassay (Biolegend Legendplex). IFN-y data are representative of three monocyte donors, IL-1 is representative of two of three donors (Figure 20C). Statistical analysis was performed using paired, parametric Student's t-tests, with significance levels denoted as follows: ** p < 0.01 , *** p < 0.001. **** p < 0.0001.

[0513] To determine their T cell activation potential, macrophages differentiated in the presence of KB16A.41.2 were used in a mixed leukocyte reaction (MLR) with naive T cells. Macrophages differentiated in the presence of KB16A.41.2 were able to elicit better T cell survival (assessed by enumerating T cells at the end of culture), increased T cell activation (assessed by activation marker CD25), and improved pro-inflammatory cytokine production (assessed by secretion of IFN-y and IL- 13).

[0514] 8.2 In Vivo Studies ofKB16A.41.2

[0515] KB16A.41.2 has demonstrated anti-tumor activity in two humanized mouse xenograft models of HLA-G expressing melanoma or bladder carcinoma.

[0516] In the first proof-of-concept in vivo study, KB16A.41.2 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 ~150 mm3(Day 0). Randomized groups of mice, each containing mice from three different HuHSC donors, were intravenously (I.V.) dosed with isotype control antibody (hlgG420 mg / kg), KB16A.41.2 (hlgG420 mg / kg) or a N. benthamiana produced MK-4830 (Clinical comparator- hlgG420 mg / kg) twice a week for 24 days. Tumor volume was monitored until Day 31. The results indicate significant tumor weight reduction 31 days after treatment in mice treated with KB16A.41.2 (-24%) or the Clinical comparator (-25%) compared to animals dosed with isotype control. Additionally, KB16A.41.2 treatment increased the percent of CD33+CD11b- myeloid cells and the percentage of HLA-DR+ and CD86+ CD11 b+CD33+ cells in the tumor microenvironment compared to isotype control treated animals.

[0517] 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 grown until average tumor sizes reached ~100 mm3(Day 0). Randomized groups of mice, each containing mice from three different huHSC donors, were then intraperitoneally dosed with isotype control antibody (hlgG4 10 mg / kg), KB16A.41.2 (hlgG4 10 mg / kg), Atezolizumab (hlgG4 10mg / kg), or KB16A.41.2 and Atezolizumab in combination (10 mg / kg each) twice a week starting on Day 1 for five total doses. Tumor volume was monitored over the course of the study until Day 17. The data imply impaired tumor growth over the course of the study in mice treated with KB16A.41.2, Atezolizumab, or KB16A.41.2+Atezolizuamb as demonstrated by significant reduction in tumor growth and increased CD8+ T-cells and CD33+CD11 b-CD86+ cell subsets in the tumor microenvironment. KB16A.41.2 treatment reduced tumor volume by -30% compared to isotype control, similar to atezolizumab positive control, while concomitant treatment of KB16A and atezolizumab lead to >50% tumor burden reduction. The study was terminated early (Day 17) due to the development of graft versus host disease (GvHD) in multiple animals.

[0518] 8.2. 1 Proof-Of-Concept Efficacy in Humanized Mouse Xenograft Model Study Design

[0519] A preliminary proof-of-concept in vivo efficacy study was performed to assess KB16A.41.2 in the treatment of subcutaneous HLA-G overexpressing SK-MEL-5 melanoma in a xenograft model in female HuHSC-NOG EXL mice. A human lgG4 isotype antibody produced by Leinco Technologies (LT9020) was used as a negative control. In this model, immunodeficient NOG-EXL (NOD.Cg- Prkdcscidll2rgtm1Su9Tg(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, >25% 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 nine mice per group (three mice from each human CD34+ donor cohort per treatment group) when mean tumor size was 150 mm3after approximately 29 days, (Day 0 tumor randomization). Tumor-bearing mice were injected I.V. with 20 mg / kg of KB16A.41.2, a Clinical Comparator, or a hlgG4 isotype control mAb twice a week for 24 days (eight total doses) following randomization. Tumor volumes were monitored twice weekly following the initiation of treatment.

[0520] Table 7 Proof-of-Concept Mouse Xenograft Efficacy Study Design

[0521] ''N. benthamiana produced MK-4830; a hlgG4 anti-LILRB2 mAb

[0522] 2Animals were dosed on Days 0, 3, 7, 10, 14, 17, 21, and 24 (8 total doses) Tumor Cell Preparation

[0523] HLA-G overexpressing SK-MEL-5 cells were maintained in vitro in MEM medium supplemented with 10% fetal bovine serum, 0.01mM NEAA at 37°C in an atmosphere of 5% CO2 in air. Cells were seeded and propagated until cells reached 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.

[0524] Tumor Implantation

[0525] Each mouse was inoculated subcutaneously in the right upper flank region with HLA-G overexpressing SK-MEL-5 cells (5 x 105) in 0.2 mL of PBS mixed with Matrigel (1 :1) for tumor development. Randomization started when the mean tumor size reached 146.55 mm3. The date of randomization was denoted as day 0; treatment was initiated on the same day of randomization (Day 0).

[0526] Tumor Measurement

[0527] 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.

[0528] 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.

[0529] 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:

[0530] • Emaciated - skeletal structure extremely prominent, little or no flesh cover, vertebrae distinctly segmented.

[0531] • Persistent hypothermia.

[0532] • Blood stained or mucopurulent discharge from any orifice.

[0533] • Labored breathing, particularly accompanied by nasal discharge or cyanosis.

[0534] • Hind limb paralysis or weakness (cannot reach food and water).

[0535] • Severe anemia, mainly indicated by pale feet or ear.

[0536] • Severe infection.

[0537] • Incontinence or diarrhea over 48 hours or severe dehydration.

[0538] •

[0539] Randomization

[0540] The randomization started when the mean tumor size reached 146.55 mm3. All animals were randomly allocated to study groups, nine mice in each group. Randomization was performed based on “Matched distribution” method.

[0541] The date of randomization was denoted as Day 0.

[0542] Study Termination

[0543] The study was terminated on Day 31.

[0544] Test and Control Articles

[0545] Table 8 Test and Control Articles 8.2.2 Results - Efficacy Outcomes - Tumor Volume Growth

[0546] The treatment (20 mg / kg) was performed twice a week (BIW) for 24 days. Tumor volumes (Figure 21)Error! Reference source not found, were also measured twice per week after randomization until termination on Day 31. Day 0 started when the mean tumor size reached 146.55 mm3and mice were randomized into groups of nine with three mice per HSC CD34+ donor per group (n=9). As shown in Figure 14, the mean tumor weights at Day 31 were significantly reduced in KB16A.41.2 and Clinical Comparator treated animals compared to Isotype control. On Day 31 , the mean tumor volume in the KB16A.41.2 or Clinical Comparator Group was treatment group was 16.5% (129.1 mm3) or 14.99% (117.2mm3) less than Isotype control group, respectively (Table 9).

[0547] 8.2.3 Tumor Growth Inhibition

[0548] Tumor Growth Inhibition (TGI) and Tumor volume of the treatment - control groups (T / C) calculation based on the tumor size data on Day 31 are presented in Table 9.

[0549] Table 9 TGI and T / C calculation on Day 31

[0550] SK-MEL-5 Tumor Growth Inhibition; ATumor Volume = Vt -VO; VO is the individual’s tumor volume on the day of randomization (day 0), Vt is its tumor volume after the day of randomization (Day 1-31) on a given day (day t). TGI% = (1-Ti / Ci) * 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

[0551] 8.2.4 Tumor Weights

[0552] At the end of study on Day 31 , tumors were collected and weighed. The mean tumor weights are graphed in Figure 22.

[0553] 8.2.5 Flow Cytometry Analysis of Tumor Immune Cells

[0554] 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. Results are shown in Figure 23.

[0555] 8.2.6 Flow Cytometry Analysis of Human CD45+ Immune Cells in Blood, Spleens, and Tumors

[0556] 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.

[0557] Results are shown in Figure 24.

[0558] 8.2.7 Analysis of HLA-G Protein Expression in Tumors by Western Blot

[0559] Tumor samples were harvested on Day 31 and analyzed via Western Blot assay to determine if hHLA-G expression was maintained in all tumors. As presented in Figure 25, Error! Reference source not found. hHLA-G was expressed in all tumors.

[0560] 8.2.8 Summary

[0561] In this first proof-of-concept in vivo study, the therapeutic efficacy of KB16A.41.2 was investigated in a SK-MEL-5 (melanoma) cell line derived subcutaneous xenograft model in HuHSC-NOG-EXL mice.

[0562] Compared to vehicle group, no statistically significant reduction in tumor volumes were detected in any treatment groups as determined by in-life tumor volume measurements. Single agent KB16A.41.2 and the Clinical Comparator (N. benth produced MK-4830) had TGIs 16.52% and 14.99% respectively, there was no significant difference when compared with the hlgG4 isotype (p>0.05) at Day 31.

[0563] In contrast, KB16A.41.2 and the Clinical Comparator treated groups on Day 31 had statistically significant reduced tumor burden as determined by whole tumor weights. Treatment groups KB16A.41.2 and the Clinical Comparator had whole tumor weights (mg) of 843.86 (p<0.05) and 826.28 (p<0.05) respectively, vs 1108.03 mg in the hlgG4 isotype group.

[0564] Flow cytometry analysis of tumors on Day 31 revealed that treatment groups KB16A.41.2 and the Clinical Comparator had no significant impact on human CD45+, human CD3+, or human CD4+ immune cells reconstruction rate in blood, spleen and tumor samples comparing with the hlgG4 isotype. However, there were significant reductions in CD8+ immune cells in the tumors of the Clinical comparator treated animals and in CD8+ HLA-DR+ immune cells in the spleens of KB16A.41.2 treated animals. For myeloid cells, absolute counts of CD33+CD11b+ subsets were decreased in KB16A.41.2 and the Clinical Comparator treated groups in both tumor and spleen samples, however, the H LA-DR and CD86 expression on this subset was significantly increased in tumors of KB16A.41.2 and / or the Clinical Comparator treatment groups when compared to Isotype control. Additionally, the percent of CD33+CD11b- cells were increased in KB16A.41.2 and the Clinical Comparator treated groups in both tumor and spleen samples.

[0565] In summary, evidence of anti-tumor activity was observed in this study with KB16A.41.2 and the Clinical Comparator demonstrated by significant reduction in tumor weight and increased myeloid cell activation in the tumor microenvironment. Additional studies should be performed to fully understand the efficacy and safety of these anti-LILRB2 mAbs in vivo.

[0566] 8.3 Confirmatory Efficacy in Humanized Mouse Xenograft Model

[0567] Study Design

[0568] A follow-on confirmatory in vivo efficacy study was performed to assess KB16A.41.2 in the treatment of subcutaneous HLA-G expressing HT-1376 bladder cancer cells in a xenograft model in female HuHSC-NOG EXL mice (Table 10). A human lgG1 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.

[0569] Table 10 Confirmatory Mouse Xenograft Efficacy Study Design

[0570] 1Animals were dosed on Days 1 , 4, 8, 11, and 15

[0571] Tumor Cell Preparation

[0572] 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. 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.

[0573] Tumor Implantation

[0574] 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. Animals were undisturbed for up to seven days before observing for tumor growth.

[0575] Tumor Measurement

[0576] 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 (100 mm3), tumors and body weights were measured at least two times per week for the duration of the study. One individual was responsible for tumor measurements for the duration of the study.

[0577] Randomization and Dose Selection

[0578] When the average tumor volume reached 100 mm3, mice were randomly assigned to the respective treatment groups and dosed within 24 hours of randomization.

[0579] Reference Day: Day 0 is defined by the day of randomization.

[0580] Study Termination

[0581] The study was terminated 17 days after randomization (Day 17).

[0582] Test and Control Articles

[0583] Table 11 Test and Control Articles

[0584] Results

[0585] 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 12).

[0586] Table 12 Average Tumor Volume at Randomization (Day 0)

[0587] Efficacy Outcomes - Tumor Volume

[0588] The tumor volume growth curves between randomization grouping and study termination are shown in Figure 26, Figure 27, and described Table 13 below.

[0589] Table 13 TGI and T / C calculation on Day 17

[0590] HT-1376 Tumor Growth Inhibition; TGI (%) = (1-Ti / Ci) * 100; Ti and Ci are the mean tumor volumes of the treatment and control groups, respectively, on a given day.

[0591] Flow Cytometry Analysis of Tumor Immune Cells

[0592] At termination (Day 17), all tumor samples were processed as single cell suspensions and stained for the following panel of markers: Live / Dead Dye, mCD45, hCD45, hCD19, h,CD56, hCD3, hCD4, hCD8, hHLA-DR, hCD33, hCD11b, 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.

[0593] Flow Cytometry Analysis of Human Blood CD45+lmmune Cells

[0594] At termination, all blood samples were stained as per 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.

[0595] Results are shown in Figure 28.

[0596] Summary

[0597] In this study, the therapeutic efficacy of KB16A.41.2 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).

[0598] 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 KB16A.41.2, Atezolizumab, and KB16A.41.2+Atezolizuamb, the TGI compared to the hlgG1 isotype control group was 28.78%%, 27.18%, and 54.63%, respectively. In the KB16A.41.2 and Atezolizumab groups, maximum TGI was observed on Day 3 in Atezolizuamb treated mice and on Day 10 in KB16A.42.1 treated mice with a TGI compared to the hlgG1 isotype control group of 32.71 % and 37.67%, respectively.

[0599] Flow cytometry analysis of tumors on Day 17 revealed that treatment groups KB16A.41.2+Atezolizuamb and / or Atezolizumab had significantly increased number of human CD3+, CD8+ and CD4+ T cells within the tumor microenvironment compared to Isotype control group. In tumor T-cells subsets, the absolute number of CD4+HLA-DR+, CD4+CD69+, and CD4+PD-1+ T-cells were significantly increased in the Atezolizuamb treated mice. Additionally, the absolute number of CD8+ cells positive for HLA-DR, CD69 and PD-1 was significantly increased in KB16A.41.2+Atezolizumab and / or Atezolizumab treated groups compared to Isotype control.

[0600] For myeloid cells, the percent of CD11b+ and CD33+CD11b- cells were significantly decreased in KB16A.41.2+Atezolizumab and / or Atezolizumab treated groups compared to Isotype control. In myeloid cell subsets in HLA-DR+ CD33+CD11 b+ cells were significantly increased in Atezolizumab treated mice, as well as the percent of CD86+ CD33+CD11b- cells.

[0601] Animals experienced mild GvHD symptoms in all study Groups during the study which resulted in the duration of the study being shortened. The termination day was determined due to GvHD progression and set at Study Day 17 to maintain group sizes at > n=8 in all treatment groups.

[0602] In summary, 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 KB16A.42.1 , Atezolizumab, or KB16A.42.1+Atezolizuamb treatment as demonstrated by significant reduction in tumor growth and increased immune cells in the tumor microenvironment. Furthermore, this study revealed synergistic anti-tumor effects of KB16A.42.1 and Atezolizumab when dosed in combination.

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: 39;(b) a CDRH2 selected from the group consisting of SEQ ID NO 40 to SEQ ID NO 80; and(c) a CDRH3 selected from the group consisting of SEQ ID NO 81 to SEQ ID NO 130; and a light chain variable region comprising:(d) a CDRL1 selected from the group consisting of SEQ ID NO: 131 to SEQ ID NO: 164;(e) a CDRL2 selected from the group consisting of SEQ ID NO: 165 to SEQ ID NO: 184; and(f) a CDRL3 selected from the group consisting of SEQ ID NO: 185 to SEQ ID NO: 228.

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: 1 , 40 and 81;(b) CDRHs according to SEQ ID NO: 2, 41 and 82;(c) CDRHs according to SEQ ID NO: 3, 42 and 83;(d) CDRHs according to SEQ ID NO: 7, 45 and 87;(e) CDRHs according to SEQ ID NO: 12, 50 and 93;(f) CDRHs according to SEQ ID NO: 4, 43 and 84;(g) CDRHs according to SEQ ID NO: 8, 50 and 88;(h) CDRHs according to SEQ ID NO: 7, 46 and 89;(i) CDRHs according to SEQ ID NO: 25, 65 and 113;(j) CDRHs according to SEQ ID NO: 36, 78 and 127;(k) CDRHs according to SEQ ID NO: 6, 44 and 86;(l) CDRHs according to SEQ ID NO: 30, 73 and 121 ;(m) CDRHs according to SEQ ID NO: 38, 47 and 129;(n) CDRHs according to SEQ ID NO: 7, 72 and 120;(o) CDRHs according to SEQ ID NO: 14, 53 and 96; or(p) CDRHs according to SEQ ID NO: 5, 57, 102.

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: 131 , 165 and 185;(b) CDRLs according to SEQ ID NO: 132, 166 and 186;(c) CDRLs according to SEQ ID NO: 133, 165 and 187;(d) CDRLs according to SEQ ID NO: 137, 168 and 191 ;(e) CDRLs according to SEQ ID NO: 136, 167 and 197;(f) CDRLs according to SEQ ID NO: 134, 166 and 188;(g) CDRLs according to SEQ ID NO: 133, 169 and 192;(h) CDRLs according to SEQ ID NO: 138, 170 and 193;(i) CDRLs according to SEQ ID NO: 155, 179 and 203;(j) CDRLs according to SEQ ID NO: 161 , 168 and 226;(k) CDRLs according to SEQ ID NO: 133, 168 and 190;(l) CDRLs according to SEQ ID NO: 164, 168 and 221 ;(m) CDRLs according to SEQ ID NO: 163, 168 and 227;(n) CDRLs according to SEQ ID NO: 164, 179 and 220;(o) CDRLs according to SEQ ID NO: 142, 172 and 200; or(p) CDRLs according to SEQ ID NO: 164, 165 and 206.

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, 40 and 81 and CDRLs according to SEQ ID NO: 131 , 165 and 185.

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: 229, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 279.

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, 41 and 82 and CDRLs according to SEQ ID NO: 132, 166 and 186.

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: 230, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 280.

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, 42 and 83 and CDRLs according to SEQ ID NO: 133, 165 and 187.

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: 231, and the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 281.

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: 7, 45 and 87 and CDRLs according to SEQ ID NO: 137, 168 and 191.

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: 12, 50 and 93 and CDRLs according to SEQ ID NO: 136, 167 and 197.

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: 4, 43 and 84 and CDRLs according to SEQ ID NO: 134, 166 and 188.

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: 8, 50 and 88 and CDRLs according to SEQ ID NO: 133, 169 and 192.

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: 7, 46 and 89 and CDRLs according to SEQ ID NO: 138, 170 and 193.

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: 25, 65 and 113 and CDRLs according to SEQ ID NO: 155, 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: 36, 78 and 127 and CDRLs according to SEQ ID NO: 161 , 168 and 226.

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: 6, 44 and 86 and CDRLs according to SEQ ID NO: 133, 168 and 190.

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: 30, 73 and 121 and CDRLs according to SEQ ID NO: 164, 168 and 221.

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: 38, 47 and 129 and CDRLs according to SEQ ID NO: 163, 168 and 227.

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: 7, 72 and 120 and CDRLs according to SEQ ID NO: 164, 179 and 220.

21. 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: 14, 53 and 96 and CDRLs according to SEQ ID NO: 142, 172 and 200.

22. 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: 5, 57, 102 and CDRLs according to SEQ ID NO: 164, 165 and 206.

23. The antibody, or antigen-binding fragment thereof, of any one of the preceding claims, wherein the antibody binds to LILRB2.

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