Pharmaceutical formulations and methods of use of anti-ILT4 antibodies or antigen-binding fragments thereof

Abstract

The present invention relates to stable formulations of antibodies or antigen-binding fragments thereof that bind to human immunoglobulin-like transcript 4 (ILT4). Also provided are methods of treating various cancers using the formulations disclosed herein.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 336,670, filed April 29, 2022, the disclosure of which is incorporated herein in its entirety.

[0002] REFERENCE TO ELECTRONICALLY SUBMITTED SEQUENCE LISTING This application contains a Sequence Listing that has been submitted electronically in XML format and is incorporated by reference in its entirety. The XML file was created on Mar. 21, 2023, has the filename 25401_WO_PCT_SL.XML, and is 131,718 bytes in size.

[0003] The present invention relates to pharmaceutical formulations of antibodies or antigen-binding fragments thereof that bind to human interleukin-like transcript 4 (ILT4) and their use in the treatment of various diseases (eg, cancer). [Background technology]

[0004] A common strategy used by tumor cells to evade innate and adaptive immune responses is associated with aberrant expression of human leukocyte antigen (HLA)-G (Curigliano et al. Clin Cancer Res. 2013; 19(20): 5564-5571 and Gonzalez et al. Crit Rev Clin Lab Sci. 2012; 49(3): 63-84). HLA-G can directly inhibit immune cell function through receptor binding and / or through impaired trogocytosis and chemotaxis (Morandi et al. Cytokine Growth Factor Review. 2014, 25: 327-335 and Lin et al. Mol Med. 2015, 21: 782-791). For example, high expression of HLA-G in multiple types of tumors, including colorectal, pancreatic, endometrial, lung, breast, ovarian and gastric cancer, is associated with advanced disease stage, tumor invasiveness, metastatic potential and poor prognosis (Lin et al. Mol Med. 2015, 21: 782-791; and Loumange et al. Int J Cancer. 2014, 135, 222: 581-597). Antibody-mediated blockade of HLA-G function in transgenic mouse models has been shown to inhibit tumor development and block myeloid-derived suppressor cell (MDSC) expansion (Loumange et al. Int J Cancer. 2014, 135, 222: 581-597; Lin et al. Hum Immunol. 2013, 74: 439-446; and Agaugue et al. Blood. 2011, 117: 7021-7031). Binding of HLA-G to ILT4 can directly inhibit the function of monocytes, dendritic cells, and neutrophils, thus potentially compromising the innate immune antitumor response.Interaction of HLA-G with monocytes through ILT4 inhibits maturation of human monocyte-derived antigen-presenting cells (APCs), resulting in reduced expression of MHC class II antigens and costimulatory molecules through activation of Stat3 (Colonna et al. J Immunol. 1998, 160: 3096-3100; Allan et al. J Exp Med. 1999, 189(7): 1149-1155; and Liang et al. Proc Natl Sci USA. 2008, 105(24): 8357). Using human monocyte-derived dendritic cells (DCs) and ILT4 transgenic mice, it was shown that HLA-G inhibits the maturation / activation of myeloid DCs and induces the generation of tolerogenic APCs, and the induction of tolerogenic DCs by HLA-G was mediated through the disruption of the MHC class II presentation pathway (Ristich et al. Eur J Immunol. 2005, 35: 1133-1142). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Curigliano et al. Clin Cancer Res. 2013; 19(20): 5564-5571 [Non-Patent Document 2] Gonzalez et al. Crit Rev Clin Lab Sci. 2012; 49(3): 63-84 [Non-Patent Document 3] Morandi et al. Cytokine Growth Factor Review. 2014, 25: 327-335 [Non-Patent Document 4] Lin et al. Mol Med. 2015, 21: 782-791 [Non-Patent Document 5] Loumange et al. Int J Cancer. 2014, 135, 222: 581-597 [Non-Patent Document 6] Lin et al. Hum Immunol. 2013, 74: 439-446 [Non-Patent Document 7] Agaugue et al. Blood. 2011, 117: 7021-7031 [Non-Patent Document 8] Colonna et al. J Immunol. 1998, 160: 3096-3100 [Non-Patent Document 9] Allan et al. J Exp Med. 1999, 189(7): 1149-1155 [Non-Patent Document 10] Liang et al. Proc Natl Sci USA. 2008, 105(24): 8357 [Non-Patent Document 11] Ristich et al. Eur J Immunol. 2005, 35: 1133-1142 Summary of the Invention [Problem to be solved by the invention]

[0006] There is an unmet medical need for patients suffering from cancers (e.g., myeloid "rich" tumors) that do not respond to T cell therapy but may benefit from alleviation of tumor tolerance mediated by tissue associated macrophages / MDSCs. ILT4 blockade therapy would fill this need and would be differentiated from current T cell-targeting antibodies (e.g., anti-PD1, anti-TIGIT) by alleviating the suppression of tolerogenic myeloid cells in the tumor microenvironment. [Means for solving the problem]

[0007] In one aspect, provided herein is a formulation of an anti-human ILT4 (anti-ILT4) antibody or antigen-binding fragment thereof, wherein the formulation comprises: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM of a buffer; (iii) about 6% to about 8% weight / volume (w / v) of a non-reducing sugar; (iv) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (v) about 1 mM to about 20 mM of an antioxidant; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the following structure: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (sequence and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); wherein the buffer is an L-histidine buffer, an acetate buffer or a citrate buffer, wherein the non-reducing sugar is a disaccharide, wherein the non-ionic surfactant is polysorbate 20 or polysorbate 80, and wherein the antioxidant is methionine.

[0008] In certain embodiments, the buffer is selected from the group consisting of an L-histidine buffer, an acetate buffer, and a citrate buffer. In one embodiment, the buffer is an L-histidine buffer. In another embodiment, the buffer is an acetate buffer. In yet another embodiment, the buffer is a citrate buffer.

[0009] In some embodiments, the non-reducing sugar is sucrose.

[0010] In certain embodiments, the non-ionic surfactant is polysorbate 80 (PS-80) or polysorbate 20 (PS-20). In one embodiment, the non-ionic surfactant is PS-80. In another embodiment, the non-ionic surfactant is PS-20.

[0011] In some embodiments, the antioxidant is L-methionine.

[0012] Accordingly, in another aspect, provided herein is a formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM L-histidine buffer; (iii) about 6% to about 8% (w / v) sucrose; (iv) about 0.01% to about 0.10% (w / v) PS-80; and (v) about 1 mM to about 20 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof is a CDR-H1:G a heavy chain variable domain comprising: YYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0013] In some embodiments, the formulation comprises about 8 mM to about 12 mM L-histidine buffer.

[0014] In certain embodiments, the formulation comprises about 5 mM to about 10 mM L-methionine.

[0015] In another embodiment, the formulation comprises about 0.01% to about 0.05% (w / v) PS-80.

[0016] In yet another embodiment, the formulation comprises about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 15 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL, or about 150 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 12.5 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 15 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 25 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 50 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 75 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 100 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 125 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 150 mg / mL. In a particular embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody.

[0017] Thus, in one particular embodiment, the formulation comprises about 25 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0018] In another specific embodiment, the formulation comprises about 50 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0019] In yet another specific embodiment, the formulation comprises about 75 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0020] In yet another specific embodiment, the formulation comprises about 100 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0021] In yet another specific embodiment, the formulation comprises about 125 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0022] In certain embodiments of the various formulations provided herein, the formulation has a pH range of about 5.0 to about 6.8. In some embodiments, the formulation has a pH range of about 5.5 to about 6.0. In other embodiments, the formulation has a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.0. In one embodiment, the formulation has a pH of about 5.5. In another embodiment, the formulation has a pH of about 5.6. In yet another embodiment, the formulation has a pH of about 5.7. In yet another embodiment, the formulation has a pH of about 5.8. In another embodiment, the formulation has a pH of about 5.9. In yet another embodiment, the formulation has a pH of about 6.0.

[0023] Thus, in one particular embodiment, the pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprises: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer, pH about 5.5; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) polysorbate 80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16). ), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0024] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0025] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0026] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0027] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0028] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0029] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58.

[0030] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0031] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains comprising the amino acid sequence set forth in SEQ ID NO:2 and two light chains comprising the amino acid sequence set forth in SEQ ID NO:7.

[0032] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains consisting of the amino acid sequences set forth in SEQ ID NO:2 and two light chains consisting of the amino acid sequences set forth in SEQ ID NO:7.

[0033] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0034] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains comprising the amino acid sequence set forth in SEQ ID NO:80 and two light chains comprising the amino acid sequence set forth in SEQ ID NO:7.

[0035] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody comprising two heavy chains consisting of the amino acid sequences set forth in SEQ ID NO:80 and two light chains consisting of the amino acid sequences set forth in SEQ ID NO:7.

[0036] In one embodiment, the formulation is a liquid formulation. In one embodiment, the formulation is a frozen formulation. In another embodiment, the liquid formulation is stored at about 5° C. In yet another embodiment, the formulation is stored frozen at about −20° C. or below. In yet another embodiment, the formulation is stored frozen at about −70° C. or below. In yet another embodiment, the liquid formulation is a solution reconstituted from a lyophilized formulation.

[0037] In certain embodiments, after storage of the formulation at about 3° C. to about 5° C. (e.g., 3° ​​C., 4° C., 5° C.) for up to 6 months, (i) the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) the OD 350-500 (iii) the turbidity of the formulation is at most about 0.135 as measured by high performance ion exchange chromatography; (iv) the number of subvisible particles of particles at least 2 μm in size is at most about 3500 as measured by microflow imaging; and / or (v) the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping.

[0038] In some embodiments, the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% when measured by ultra performance size exclusion chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months.

[0039] In some embodiments, the formulation is stored at about 3° C. to about 5° C. for 6 months and then the OD 350-500 When measured at 100° C., the turbidity of the formulation is a maximum of about 0.135.

[0040] In some embodiments, the % major peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the % major peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography.

[0041] In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the number of sub-visible particles having a size of at least 2 μm is up to about 3500 as measured by microflow imaging.

[0042] In some embodiments, the formulation is stored at about 3° C. to about 5° C. for 6 months and the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping. In one embodiment, the amino acid residue is W7 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In another embodiment, the amino acid residue is W102 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In yet another embodiment, the amino acid residue is M253 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In yet another embodiment, the amino acid residue is M359 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80. In yet another embodiment, the amino acid residue is M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80.

[0043] In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of one amino acid residue selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of two amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of three amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of four amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, the % oxidation of all five amino acid residues from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3°C ​​to about 5°C for 6 months.

[0044] In yet another aspect, there is provided herein a method of treating cancer in a human patient in need of such treatment, wherein the method comprises administering a therapeutically effective amount of a pharmaceutical formulation described herein.

[0045] In yet another aspect, there is provided herein a use of a pharmaceutical formulation described herein for the preparation of a medicament for treating cancer. [Brief description of the drawings]

[0046] [Figure 1A] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1B] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1C] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1D] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1E] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1F] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1G] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1H] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 1I] 1A-1I show early stability studies of various anti-ILT4 formulations, including turbidity (OD350-500) (FIGS. 1A-1C), % HMW (FIGS. 1D-1F) or % monomer (FIGS. 1G-1I) at storage conditions of 5° C. (FIGS. 1A, 1D, 1G), 25° C. (FIGS. 1B, 1E, 1H) or 40° C. (FIGS. 1C, 1F, 1I), as described in Example 4. [Figure 2A] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2B]2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2C] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2D] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2E] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2F] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2G]2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2H] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 2I] 2A-2I show early stability studies of various anti-ILT4 formulations including % acidic variant (FIGS. 2A-2C), % main peak (FIGS. 2D-2F) or % basic variant (FIGS. 2G-2I) at storage conditions of 5° C. (FIGS. 2A, 2D, 2G), 25° C. (FIGS. 2B, 2E, 2H) or 40° C. (FIGS. 2C, 2F, 2I), as described in Example 4. [Figure 3A] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3B] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3C]3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3D] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3E] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3F] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3G] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3H]3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 3I] 3A-3I show pH ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIGS. 3A-3C), % HMW (FIGS. 3D-3F) or % monomer (FIGS. 3G-3I) at storage conditions of 5° C. (FIGS. 3A, 3D, 3G), 25° C. (FIGS. 3B, 3E, 3H) or 40° C. (FIGS. 3C, 3F, 3I), as described in Example 5. [Figure 4A] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4B] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4C] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4D]4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4E] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4F] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4G] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4H] 4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 4I]4A-4I show pH ranging stability studies of various anti-ILT4 formulations including % acidic variant (FIG. 4A-4C), % main peak (FIG. 4D-4F) or % basic variant (FIG. 4G-4I) at storage conditions of 5° C. (FIG. 4A, FIG. 4D, FIG. 4G), 25° C. (FIG. 4B, FIG. 4E, FIG. 4H) or 40° C. (FIG. 4C, FIG. 4F, FIG. 4I) as described in Example 5. [Figure 5A] 5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 5B] 5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 5C] 5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 5D]5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 5E] 5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 5F] 5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 5G] 5A-5G show surfactant ranging stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 5A), % HMW (FIG. 5B), % monomer (FIG. 5C), number of sub-visible particles before stirring started (FIG. 5D), number of sub-visible particles after 3 days of stirring (FIG. 5E), number of sub-visible particles after 7 days of stirring (FIG. 5F) or number of sub-visible particles after 7 days of ambient control (FIG. 5G), as described in Example 6. [Figure 6A]6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6B] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6C] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6D] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6E]6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6F] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6G] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6H] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6I]6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6J] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 6K] 6A-6K show photostress stability studies of various anti-ILT4 formulations including turbidity (OD350-500) (FIG. 6A), % HMW (FIG. 6B), % monomer (FIG. 6C), % acidic variant (FIG. 6D), % main peak (FIG. 6E), % basic variant (FIG. 6F), % oxidation of M359 (FIG. 6G), % oxidation of M253 (FIG. 6H), % oxidation of M429 (FIG. 6I), % oxidation of W102 (FIG. 6J) or % oxidation of W7 (FIG. 6K), as described in Example 7. [Figure 7] FIG. 7 shows the initial and 3-day agitation results from a surfactant blending study, as described in Example 6. [Figure 8] FIG. 8 shows the results of a 7 day agitation and a 7 day ambient control from a surfactant ranging study, as described in Example 6. [Figure 9] FIG. 9 shows the results of a light stress study using the antioxidant L-methionine, as described in Example 7. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0047] Definitions and Abbreviations As used throughout this specification and the appended claims, the following abbreviations apply: [Table 1]

[0048] Unless clearly defined elsewhere in this specification, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

[0049] As used throughout this specification and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0050] References to "or" indicate either or both possibilities, unless the context clearly dictates otherwise. In some cases, "and / or" is used to emphasize either or both possibilities.

[0051] As used herein, "acidic variant" refers to an anti-ILT4 antibody that is more acidic (e.g., as determined by cation exchange chromatography) compared to the major species of the anti-ILT4 antibody. Such acidic variants are detected by various chromatographic purification methods to separate molecular variants by charge, e.g., ion exchange chromatography, e.g., cation exchange chromatography or WCX-10 HPLC (weak cation exchange chromatography), and optionally subsequently detected by mass spectrometry. In general, acidic variants have a lower isoelectric point (pI) than the major species and may be more acidic in nature, e.g., due to methionine oxidation, sialylation of asparagine residues, or deamidation variants of the antibody, or a combination thereof. In one embodiment, the anti-ILT4 antibody acidic variant is the anti-ILT4 antibody species identified by the acidic variant peak in Figure 6D and eluted according to the cation ion exchange method described in Example 10. In the ion exchange chromatography method, "% acidic variant species" refers to the total area of ​​the acidic variant peak divided by the total area of ​​all peaks in the elution chromatogram. In one embodiment, the anti-ILT4 antibody acidic variant is identified by a peak eluting before the main peak according to a cationic ion exchange method. In another embodiment, the anti-ILT4 antibody acidic variant is identified by a peak eluting before the main peak according to a weak cationic ion exchange method. In an ion exchange chromatography method, "% acidic variant" refers to the total area of ​​the acidic species peak divided by the total area of ​​all peaks in the elution chromatogram.

[0052] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y is generally measured by the dissociation constant (K DAffinity can be measured by common methods known in the art, such as KinExA and Biacore.

[0053] As used herein, the term "antibody" includes, but is not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), fully human antibodies, and chimeric antibodies.

[0054] As used herein, unless otherwise indicated, "antigen-binding fragment" refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to bind to the antigen bound by the full-length antibody (e.g., a fragment that retains one or more CDR regions). Examples of antibody-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, and individual antibody heavy or light chains and individual heavy or light chain variable regions.

[0055] As used herein, "basic variant" refers to an anti-ILT4 antibody that is more basic (e.g., as determined by cation exchange chromatography) compared to the major species of the anti-ILT4 antibody. Such basic variants are detected by various chromatographic purification methods to separate molecular variants by charge, for example, ion exchange chromatography, for example, cation exchange chromatography (e.g., the method described in Example 10) or WCX-10 HPLC (weak cation exchange chromatography), and optionally subsequently detected by mass spectrometry. In general, basic variants have a higher pH than the major species and may be more acidic in nature due to modifications or differences from the major species. In one embodiment, the anti-ILT4 antibody basic species is identified by a peak that elutes after the major peak according to a cation ion exchange method. In another embodiment, the anti-ILT4 antibody basic species is identified by a peak that elutes after the major peak according to a weak cation ion exchange method. In an ion exchange chromatography method, "% basic species" refers to the total area of ​​the basic species peak divided by the total area of ​​all peaks in the elution chromatogram.

[0056] A "Fab fragment" is composed of one light chain and the CH1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule. A "Fab fragment" can be the product of papain cleavage of an antibody.

[0057] The "Fc" region comprises two heavy chain fragments comprising the CH1 and CH2 domains of an antibody, held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domain.

[0058] A "Fab' fragment" is a fragment that contains one light chain and one V HThe Fab' fragment comprises a portion or fragment of one heavy chain that includes the CH1 domain and the CH2 domain and also includes the region between the CH1 and CH2 domains, whereby interchain disulfide bonds can form between the two heavy chains of the two Fab' fragments to form an F(ab')2 molecule.

[0059] An "F(ab')2 fragment" contains two light chains and two heavy chains that contain a portion of the constant region between the CH1 and CH2 domains, thereby forming an interchain disulfide bond between the two heavy chains. An F(ab')2 fragment is thus composed of two Fab' fragments that are linked by disulfide bonds between the two heavy chains. An "F(ab')2 fragment" may be the product of pepsin cleavage of an antibody.

[0060] The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

[0061] "Isolated antibody" refers to a purified state, and in this context means that the molecule is substantially free of other biological molecules (e.g., nucleic acids, proteins, lipids, carbohydrates) or other substances (e.g., cell debris and growth medium). In general, the term "isolated" is not intended to imply the complete absence of such substances, or the absence of water, buffers or salts, unless such substances or water, buffers or salts are present in an amount that would substantially interfere with the experimental or therapeutic use of the binding compounds described herein.

[0062] As used herein, "major species" or "major" refers to an anti-ILT4 antibody species identified as the majority of antibody species in a mixture with one or more acidic or basic species. Such a major species is detected by various chromatographic purification methods to separate molecular variants by charge, for example, by ion exchange chromatography, for example, cation exchange chromatography (e.g., the method described in Example 10) or WCX-10 HPLC (weak cation exchange chromatography), and optionally subsequently detected by mass spectrometry. The mixture may be, for example, the result of an antibody preparation from mammalian cells and its post-translational modifications, upstream and downstream processing or storage. In one embodiment, the major species is identified as the major peak following a cation ion exchange method. In the ion exchange method, "% major" refers to the total area of ​​the major peak divided by the total area of ​​all peaks in the elution chromatogram. In one embodiment for measuring the major species, acidic or basic species, a Thermo Scientific ProPac WCX-10 column is used for the cation ion exchange method. In another embodiment, a Thermo Scientific ProPac WCX-10 column is used, where the mobile phase (A) is 24 mM MES pH 6.1 (containing 4% acetonitrile), the mobile phase (B) is 20 mM sodium phosphate, 95 mM NaCl pH 8.0 (containing 4% acetonitrile), and the column temperature is 35°C. In one embodiment, the following non-linear gradient is used: 0-0.6 min 22%-22% B; 0.6-15.0 min 22%-29% B; 15.0-30.0 min 29%-70% B; 30.0-30.5 min 70%-100% B; and 30.5-33.0 min 100%-100% B. In a further embodiment, the cation ion exchange method is described in Example 10.

[0063] The term "monoclonal antibody" as used herein refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence, except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically contain a large number of different antibodies having different amino acid sequences in the variable domains, often specific for different epitopes. The modifier "monoclonal" indicates the nature of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al. ((1975) Nature 256: 495), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The monoclonal antibodies can also be isolated from phage antibody libraries using the techniques described in, for example, Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0064] The terms "fully human antibody" or "human antibody" refer to an antibody that contains only human immunoglobulin protein sequences. A fully human antibody may contain mouse carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" refers to an antibody that contains only mouse immunoglobulin sequences. Alternatively, a fully human antibody may contain rat carbohydrate chains if produced in a rat, in a rat cell, or in a hybridoma derived from a rat cell. Similarly, a "rat antibody" refers to an antibody that contains only rat immunoglobulin sequences.

[0065] Generally, the basic "antibody" structural unit comprises a tetramer. In monospecific antibodies, each tetramer comprises two pairs of identical polypeptide chains, each pair having one "light chain" (about 25 kDa) and one "heavy chain" (about 50-70 kDa). The amino-terminal portion of each chain contains a "variable region" or "variable domain" of about 100-110 or more amino acids that are primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region that is primarily responsible for effector function.

[0066] Typically, human constant light chains are classified as kappa and lambda light chains. Furthermore, human constant heavy chains are typically classified as mu, delta, gamma, alpha or epsilon, defining the antibody's isotype as IgM, IgD, IgG, IgA and IgE, respectively. These IgG subtypes include, for example, IgG1 and IgG4. The present invention encompasses anti-ILT4 antibodies and antigen-binding fragments comprising any of these light and / or heavy constant chains.

[0067] As used herein, "variable region," "variable domain," "V region," or "V chain" refers to the segment of an IgG chain that is variable in sequence among different antibodies. The "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. The variable region of the heavy chain is referred to as the "V H The variable region of the light chain may be referred to as "V L". Typically, both heavy and light chain variable regions contain three hypervariable regions (also called complementarity determining regions (CDRs)) located within relatively conserved framework regions (FRs). The CDRs are usually aligned with the framework regions and enable binding to a specific epitope. Generally, from N-terminus to C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment of amino acids to each domain generally follows the definitions in "Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. No. 91-3242 (1991)," "Kabat (1978) Adv. Prot. Chem. 32:1-75," "Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616," "Chothia, et al., (1987) J Mol. Biol. 196:901-917," or "Chothia, et al., (1989) Nature 342:878-883."

[0068] "CDR" is the antibody V H One of the three hypervariable regions (H1, H2, or H3) within the non-framework regions of the β-sheet framework, or LIt indicates one of three hypervariable regions (L1, L2, or L3) in the non-framework regions of the β-sheet framework. Thus, CDRs are variable region sequences interspersed within the framework region sequences. CDR regions are well known to those skilled in the art and are defined, for example, by Kabat as the most hypervariable regions in antibody variable domains. CDR region sequences are also structurally defined by Chothia as residues that are not part of the conserved β-sheet framework and therefore can adapt to various conformations. Both terms are well recognized in the art. CDR region sequences are also defined by AbM, Contact, and IMGT. The positions of CDRs in canonical antibody variable regions have been determined by comparison of multiple structures (Al-Lazikani et al., 1997, J. Mol. Biol. 273:927-48; Morea et al., 2000, Methods 20:267-79). Since the number of residues in the hypervariable regions varies among antibodies, additional residues relative to the canonical position are conventionally numbered a, b, c, etc., next to the residue number in the canonical variable region numbering scheme (Al-Lazikani et al., supra). Such nomenclature is likewise familiar to those of skill in the art. Correspondences between numbering systems, including, for example, Kabat numbering and the IMGT specific numbering system, are familiar to those of skill in the art and are shown in Table 1 below. In some embodiments, the CDRs are defined by the Kabat numbering system. In other embodiments, the CDRs are defined by the IMGT numbering system. In yet other embodiments, the CDRs are defined by the AbM numbering system. In yet other embodiments, the CDRs are defined by the Chothia numbering system. In yet other embodiments, the CDRs are defined by the Contact numbering system.

[0069] [Table 2]

[0070] Sequence identity indicates the degree to which the amino acids of two polypeptides are identical in equal positions when the two sequences are optimally aligned.

[0071] Sequence similarity includes identical residues and non-identical amino acids that are biochemically related. Biochemically related amino acids that share similar properties and may be interchangeable are discussed above.

[0072] "Conservatively modified variants" or "conservative substitutions" refer to the substitution of amino acids in a protein with other amino acids having similar properties (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, etc.), such that such changes can often be made without altering the biological activity of the protein. Those skilled in the art will generally recognize that single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to destroy biological activity. Exemplary conservative substitutions are shown in Table 2 below.

[0073] [Table 3]

[0074] As used herein, the term "epitope" refers to the area or region on an antigen to which an antibody or antigen-binding fragment binds. Binding of an antibody or antigen-binding fragment thereof disclosed herein to an epitope means that the antibody or antigen-binding fragment thereof binds to one or more amino acid residues within the epitope.

[0075] As used herein, "treat" or "treating" a cancer or infectious condition means administering a formulation of the invention to a subject having an immune or cancerous condition or diagnosed with cancer or a pathogenic infection (e.g., viral, bacterial, fungal) to achieve at least one positive therapeutic effect (e.g., a reduction in the number of cancer cells, a reduction in tumor size, a reduction in the rate of cancer cell invasion into peripheral organs, or a reduction in the rate of tumor metastasis or growth, etc.). "Treatment" may include one or more of the following: inducing / increasing an anti-tumor immune response, stimulating an immune response to pathogens, toxins and / or self-antigens, stimulating an immune response to viral infection, reducing the number of one or more tumor markers, inhibiting tumor cell proliferation or survival, eliminating or reducing the size of one or more cancerous lesions or tumors, reducing the level of one or more tumor markers, ameliorating cancer, reducing the severity or duration of cancer, prolonging patient survival compared to the expected survival of a similar untreated patient.

[0076] The terms "cancer", "cancerous" or "malignant" refer to or describe a physiological condition in mammals that is typically characterized by unregulated proliferation of cells. Examples of cancer include, but are not limited to, carcinoma, lymphoma, leukemia, blastoma and sarcoma. More specific examples of such cancers include squamous cell carcinoma, myeloma, small cell lung cancer, non-small cell lung cancer, glioma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, gastrointestinal (ductal) cancer, renal cancer, ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colon cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, brain tumor, stomach cancer, bladder cancer, liver cancer, breast cancer, colon cancer and head and neck cancer.

[0077] The term "patient" (alternatively referred to herein as "subject" or "individual") refers to a mammal (e.g., rat, mouse, dog, cat, rabbit) that can be treated with the formulations of the invention, and most preferably a human. In some embodiments, the patient is an adult patient. In other embodiments, the patient is a pediatric patient. A patient "in need of treatment" is an individual who has been diagnosed with, is suspected of being, or is susceptible to a disease or disorder that the formulations of the invention are intended to treat or ameliorate (e.g., an ILT4-associated disease, e.g., cancer), or in whom it is desired to prevent such a disorder.

[0078] The term "therapeutically effective amount" or "effective amount" refers to the amount of a therapeutic composition or formulation introduced into a patient that is sufficient to treat a disease or condition. One of skill in the art will recognize that this level may vary depending on patient characteristics such as age, weight, etc.

[0079] The term "about," when modifying an amount of a substance or composition (e.g., mM or M), a proportion of a formulation component (v / v or w / v), the pH of a solution / formulation, or the value of a parameter characterizing a step of a method, refers to a variation in a numerical quantity that may occur, for example, through typical measuring, handling, and sampling procedures associated with preparing, characterizing, and / or using a substance or composition; through instrumental errors in these procedures; through differences in the manufacture, source, or purity of ingredients employed in making or using a composition or performing a procedure; etc. In certain embodiments, "about" may mean a variation of ±0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, or 10%.

[0080] As used herein, "x% (w / v)" corresponds to x g / 100 mL (e.g., 5% w / v corresponds to 50 mg / mL).

[0081] As used throughout this specification and claims, the phrase "consists essentially of" or variations such as "consist essentially of" or "consisting essentially of" refers to the inclusion of any recited element or group of elements, and optionally the inclusion of other elements of a similar or different nature to the recited elements, that do not substantially alter the basic or novel characteristics of the specified dosing regimen, method, or composition. As a non-limiting example, a binding compound consisting essentially of a recited amino acid sequence may contain one or more amino acids that include substitutions of one or more amino acid residues that do not substantially affect the properties of the binding compound.

[0082] The word "comprising" or variations thereof, such as "comprise" or "comprises," are used throughout this specification and claims in their inclusive sense, i.e., unless the context requires otherwise, either by express language or by necessary implication, to specify the presence of stated features but not to exclude the presence or addition of further features that may materially enhance the practice or utility of any of the embodiments of the invention.

[0083] The term "buffering agent" includes agents that maintain the pH of a solution of the formulation within an acceptable range, or, in the case of a lyophilized formulation of the invention, provides an acceptable pH of the solution prior to lyophilization.

[0084] The terms "lyophilization," "lyophilized," and "freeze-dried" refer to a process in which the material to be dried is first frozen and then the ice or frozen solvent is removed by sublimation under reduced pressure. Excipients can be included in the pre-lyophilized formulation to enhance the storage stability of the lyophilized product.

[0085] The term "pharmaceutical formulation" refers to a preparation in which the active ingredient is in a form that allows it to be effective and which does not contain additional ingredients that are toxic to the subject to which it is administered. The terms "formulation" and "pharmaceutical formulation" are used interchangeably throughout.

[0086] "Pharmaceutically acceptable" refers to excipients (vehicles, additives) and compositions that can be reasonably administered to a subject to provide an effective dose of the active ingredient employed, that are "generally regarded as safe," and that, for example, are physiologically tolerated when administered to humans and do not typically produce an allergic reaction or similar unpleasant reaction (e.g., stomach upset). In another embodiment, the term refers to molecular entities and compositions approved by a federal or state regulatory agency or listed in the United States Pharmacopeia or other generally recognized pharmacopoeias for use in animals (more particularly, humans).

[0087] A "reconstituted" formulation is one in which a lyophilized protein formulation has been prepared by dissolving the lyophilized protein formulation in a diluent such that the protein is dispersed in the reconstituted formulation. The reconstituted formulation is suitable for administration (e.g., parenteral administration), and optionally, for subcutaneous administration.

[0088] A "stable" formulation is one in which the protein therein essentially retains its physical and / or chemical stability and / or biological activity under storage or stress conditions. Various analytical techniques for measuring protein stability are available in the art and are reviewed in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability may be measured at a selected temperature for a selected period of time. For example, in one embodiment, a stable formulation is one in which no significant change is observed at refrigerated temperatures (2-8°C) for at least 6 months. In one embodiment, a stable formulation is one in which no significant change is observed at refrigerated temperatures (2-8°C) for at least 12 months. In another embodiment, a stable formulation is one in which no significant change is observed at refrigerated temperatures (2-8°C) for at least 24 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 3 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 6 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 12 months. In another embodiment, a stable formulation is one in which no significant change is observed at room temperature (23-27°C) for at least 18 months. Stability criteria for antibody formulations are as follows. Typically, the degradation of antibody monomer is 10% or less, preferably 5% or less, as measured by SEC-HPLC. Typically, the formulation is colorless or clear to slightly opaque upon visual analysis. Typically, the concentration, pH, and osmolality of the formulation vary within ±10%. Titers are typically within 60-140%, preferably 80-120%, of a control or reference. Typically, antibody clipping observed is 10% or less, preferably 5% or less, i.e., % low molecular weight species as measured, for example, by HP-SEC.Typically, antibody aggregation observed is less than 10%, preferably less than 5%, i.e., % high molecular weight species as measured, for example, by HP-SEC.

[0089] An antibody "retains physical stability" in a pharmaceutical formulation if it does not show a significant increase in aggregation, precipitation, and / or denaturation upon visual inspection of color and / or clarity, or as measured by UV light scattering, size exclusion chromatography (SEC), and dynamic light scattering. Changes in protein conformation can be assessed by fluorescence spectroscopy, which confirms the tertiary structure of a protein, and FTIR spectroscopy, which confirms the secondary structure of a protein.

[0090] An antibody "retains chemical stability" in a pharmaceutical formulation if it does not undergo significant chemical changes. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Degradation processes that often alter the chemical structure of a protein include hydrolysis or clipping (assessed by methods such as size exclusion chromatography and SDS-PAGE), oxidation (assessed by methods such as MALDI / TOF / MS or peptide mapping in combination with mass spectrometry), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, isoaspartic acid measurement), and isomerization (assessed by measurement of isoaspartic acid content, peptide mapping, etc.).

[0091] An antibody is "biologically stable" in a pharmaceutical formulation if the biological activity of the antibody at a given time is within a given range of the biological activity exhibited at the time the pharmaceutical formulation was prepared. The biological activity of the antibody can be confirmed, for example, by antigen binding assays.

[0092] The term "isotonic" means that the formulation has essentially the same osmotic pressure as human blood. Isotonic formulations generally have an osmotic pressure of about 270-328 mOsm. Slightly hypotonic is 250-269 mOsm, and slightly hypertonic is 328-350 mOsm. Osmotic pressure can be measured, for example, using a vapor pressure or ice pressure osmometer.

[0093] Anti-ILT4 antibodies and antigen-binding fragments thereof The formulations disclosed herein can be used with any antibody or antigen-binding fragment thereof that binds to ILT4 (e.g., fully human antibodies). In one embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is an antagonist. In another embodiment, the anti-ILT4 antibody or antigen-binding fragment thereof is an anti-ILT4 antibody or antigen-binding fragment thereof described herein (e.g., 1E1, 2A6, 3G7, or 2C1).

[0094] In one embodiment, the formulations can be used with the anti-ILT4 antibodies and antigen-binding fragments thereof described herein that have one or more of the following properties: * binds human ILT4 at one or more amino acid residues of LYREKKSASW (SEQ ID NO:59), TRIRPEL (SEQ ID NO:60), NGQF (SEQ ID NO:61) and / or HTGRYGCQ (SEQ ID NO:62) and / or protects LYREKKSASW (SEQ ID NO:59), TRIRPEL (SEQ ID NO:60), NGQF (SEQ ID NO:61) and / or HTGRYGCQ (SEQ ID NO:62) from deuterium (e.g., DO) exchange (e.g., as confirmed by hydrogen-deuterium exchange mass spectrometry); * Binds to human ILT4 via domain 1 (see Wilcox et al. BMC Structural Biology 2:6(2002)); * binds to the human ILT4 extracellular domain or the TM form of ILT4 expressed on the cell surface (e.g., pre-B cells, Chinese hamster ovary cells, U937 cells or Jurkat JE6 cells); * Calculated pI of about 7.29 (e.g., 7.29 or 7.30); *Experimentally determined pI approx. 7.2; * Characterized by a thermogram with a Tm onset >60°C, a Tm1 of about 65.2°C and a Tm2 of about 78.8°C; * Approximately 1.7×10 -8 K of M D binds to human ILT4 (e.g., as confirmed by surface plasmon resonance, e.g., by binding of anti-ILT4 to polyhistidine-tagged human ILT4); * Ka=5.5×10 5 M -1 s -1 (e.g., as confirmed by surface plasmon resonance, e.g., by binding of anti-ILT4 to polyhistidine-tagged human ILT4); * Kd=9×10 -3 s -1 (e.g., as confirmed by surface plasmon resonance, e.g., by binding of anti-ILT4 to polyhistidine-tagged human ILT4); * Binding of HLA-G (e.g., Fc-fused HLA-G) to human ILT4 (e.g., ILT4 on murine 3A9 T cells transfected with ILT4 and expressing ILT4) is measured, for example, with an IC of about 0.25 micrograms / mL (±0.06 micrograms / mL). 50 blocking with (e.g., as confirmed by surface plasmon resonance); * blocking the binding of HLA-A, HLA-B (e.g., fluorochrome-labeled dexamers of HLA-A, e.g., HLA*A2:01, or HLA-B, e.g., HLA*B7:02) and / or HLA-F (e.g., fluorochrome-labeled tetramers of HLA-F) to human ILT4 (e.g., ILT4 on murine 3A9 T cells transfected with ILT4 and expressing ILT4) (e.g., as determined by surface plasmon resonance); * blocking the binding of LT4 (e.g., ILT4 on murine 3A9 T cells transfected with ILT4 and expressing ILT4) to ANGPTL1, ANGPTL4 and / or ANGPTL7 (e.g., biotinylated ANGPTL proteins) (e.g., as determined by surface plasmon resonance); *Does not bind to ILT2, ILT3, ILT5, LILRB5, LILRA1, LILRA2, ILT7, ILT8 and / or ILT11; * In ILT4-transfected 3A9 cells, ILT4-mediated IL2 suppression was measured with an EC 50 Reverse with ; * For example, when mast cells express ILT4 and CD200RLa and are stimulated, for example, by antibody-mediated cross-linking of CD200RLa, rescues the ILT4:HLA-G-induced inhibition of mast cell degranulation (e.g., in the presence of plate-bound HLA-G tetramers); * Enhance lipopolysaccharide (LPS)-induced expression of proinflammatory myeloid cytokines (e.g., GM-CSF and / or TNFα) from peripheral blood mononuclear cells (PBMCs); * Enhance anti-CD3-induced expression of proinflammatory myeloid cytokines (e.g., GM-CSF and / or TNFα) from peripheral blood mononuclear cells (PBMCs); * inhibiting tumor growth in humans or, for example, in other mammals, for example, mice reconstituted with human hematopoietic stem cells harboring peripheral human CD45+ immune cells (e.g., immunodeficient NSG mice) (e.g., where the tumor is a human cutaneous melanoma tumor, for example, a human cutaneous melanoma tumor derived from cell line SKMEL5); * alleviating MDSC-mediated tumor tolerance in a tumor-bearing subject (e.g., a human subject); *Does not bind to cynomolgus ILT4 and / or mouse pirB; * staining CD14+ human monocytes and / or CD11B+ human granulocytes; and / or * Binds to one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or all 10) of the human ILT4 haplotypes.

[0095] Antibody 1E1 (Q1E) heavy chain (IgG4) Heavy chain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:1; variable domains underlined; CDRs double underlined).

[0096] Heavy Chain Variable Domain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS (SEQ ID NO:63).

[0097] Antibody 1E1 (Q1E, S54A) heavy chain (IgG4) Heavy chain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHAGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:2; variable domains underlined; CDRs double underlined).

[0098] Heavy Chain Variable Domain EVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHAGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS (SEQ ID NO:57).

[0099] Antibody 1E1 heavy chain (IgG1) Heavy chain QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:44; variable domain underlined; CDRs double underlined).

[0100] Heavy Chain Variable Domain QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLPTRWVTTRYFDLWGRGTLVTVSS (sequence number: 69).

[0101] 1E1 Heavy Chain CDRs CDR-H1: GYYWS (SEQ ID NO: 16) CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) (e.g., EINHSGSTNYNPSLKS (SEQ ID NO: 47) or EINHAGSTNYNPSLKS (SEQ ID NO: 48)) CDR-H3: LPTRWVTTRYFDL (sequence number: 18).

[0102] Antibody 1E1 (Q1E) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:3; variable domains underlined; CDRs double underlined).

[0103] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (SEQ ID NO: 70).

[0104] Antibody 1E1 (Q1E, S54A) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNANRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:4; variable domains underlined; CDRs double underlined).

[0105] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNANRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 71).

[0106] Antibody 1E1 (Q1E, N53Q) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGQSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:5; variable domains underlined; CDRs double underlined).

[0107] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGQSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 72).

[0108] Antibody 1E1 (Q1E, N53E) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGESNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:6; variable domains underlined; CDRs double underlined).

[0109] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGESNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 73).

[0110] Antibody 1E1 (Q1E, N53D) light chain (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:7; variable domain underlined; CDRs double underlined).

[0111] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 58).

[0112] Antibody 1E1 light chain (lambda) Light chain QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:45; variable domain underlined; CDRs double underlined).

[0113] Light Chain Variable Domain QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSVSKSGASASLAITGLQAEDEADYYCQSFDNSLSAYVFGGGTQLTVL (Sequence number: 77).

[0114] 1E1 Light Chain CDRs CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19) CDR-L2: GX1X2NRPS; where X1 is N, Q, E or D and X2 is S or A (SEQ ID NO:20) (e.g., GNSNRPS (SEQ ID NO:49), GQSNRPS (SEQ ID NO:50), GESNRPS (SEQ ID NO:51), GDSNRPS (SEQ ID NO:52), GNANRPS (SEQ ID NO:53), GQANRPS (SEQ ID NO:54), GEANRPS (SEQ ID NO:55), or GDANRPS (SEQ ID NO:56)) CDR-L3: QSFDNSLSAYV (sequence number: 21).

[0115] 1E1 heavy and light chain CDRs or 1E1 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising 1E1 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "1E1."

[0116] Antibody 2A6(Q1E) Heavy Chain (IgG4) Heavy chain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDSSGWYKGGAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:8; variable domain underlined; CDRs double underlined).

[0117] Heavy Chain Variable Domain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDSSGWYKGGAFDIWGQGTMVTVSS (Sequence number: 64).

[0118] Antibody 2A6 (Q1E, S102A, M119L) heavy chain (IgG4) Heavy chain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDASGWYKGGAFDIWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:9; variable domains underlined; CDRs double underlined).

[0119] Heavy Chain Variable Domain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFDASGWYKGGAFDIWGQGTLVTVSS (Sequence number: 65).

[0120] Antibody 2A6 (Q1E, D101S, M119L) heavy chain (IgG4) Heavy chain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFSSSGWYKGGAFDIWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:10; variable domain underlined; CDRs double underlined).

[0121] Heavy Chain Variable Domain EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARYFSSSGWYKGGAFDIWGQGTLVTVSS (Sequence number: 66).

[0122] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO:8, 9, 10, 64, 65 or 66, where residue 1 of SEQ ID NO:8, 9, 10, 64, 65 or 66 is Q instead of E.

[0123] 2A6 Heavy Chain CDRs CDR-H1: SYAIS (SEQ ID NO: 22) CDR-H2: GIIPIFGTANYAQKFQG (SEQ ID NO: 23) CDR-H3: YFX1X2SGWYKGGAFDI; where X1 is D or S and X2 is S or A (SEQ ID NO: 24) (e.g., YFDSSGWYKGGAFDI (SEQ ID NO: 91), YFSSSGWYKGGAFDI (SEQ ID NO: 92), YFDASGWYKGGAFDI (SEQ ID NO: 93), or YFSASGWYKGGAFDI (SEQ ID NO: 94)).

[0124] Antibody 2A6 light chain (lambda) Light chain QSVLTQPSSLSASPGASASLTCTLRSGINVDTYRIHWYQQKPGSPPQYLLRYKSDSDKHQGSGVPSRFSGSKDPSANAGILLISGLQSEDEADYYCAIWYSSTWVFGGGTQLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:11; variable domain underlined; CDRs double underlined).

[0125] Light Chain Variable Domain QSVLTQPSSLSASPGASASLTCTLRSGINVDTYRIHWYQQKPGSPPQYLLRYKSDSDKHQGSGVPSRFSGSKDPSANAGILLISGLQSEDEADYYCAIWYSSTWVFGGGTQLTVL (Sequence number: 74).

[0126] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO:11 or 74, where residue 1 of SEQ ID NO:11 or 74 is E instead of Q.

[0127] 2A6 light chain CDRs CDR-L1: TLRSGINVDTYRIH (SEQ ID NO: 25) CDR-L2: YKSDSDKHQGS (SEQ ID NO: 26) CDR-L3: AIWYSSTWV (sequence number: 27).

[0128] 2A6 heavy and light chain CDRs or 2A6 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising the 2A6 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "2A6."

[0129] Antibody 3G7(Q1E) heavy chain (IgG4) Heavy chain EVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVGEWIQLWSPFDYWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:12; variable domain underlined; CDRs double underlined).

[0130] Heavy Chain Variable Domain EVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVGEWIQLWSPFDYWGQGTLVTVSS (Sequence number: 67).

[0131] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO: 12 or 67, where residue 1 of SEQ ID NO: 12 or 67 is Q instead of E.

[0132] 3G7 Heavy Chain CDR CDR-H1: SYAMH (SEQ ID NO: 28) CDR-H2: VISYDGSNKYYADSVKG (SEQ ID NO: 29) CDR-H3: VGEWIQLWSPFDY (sequence number: 30).

[0133] Antibody 3G7 light chain (kappa) Light chain DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKFLIYAASSLQSGVPSKFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPPTFGGGTKVEIK RtVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:13; variable domain underlined; CDRs double underlined).

[0134] Light Chain Variable Domain DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKFLIYAASSLQSGVPSKFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPPTFGGGTKVEIK (Sequence number: 75).

[0135] 3G7 light chain CDR CDR-L1: RASQGISSWLA (SEQ ID NO: 31) CDR-L2: AASSLQS (SEQ ID NO: 32) CDR-L3: QQYNSYPPT (sequence number: 33).

[0136] 3G7 heavy and light chain CDRs or 3G7 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising the 3G7 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "3G7."

[0137] Antibody 2C1(Q1E) Heavy Chain (IgG4) Heavy chain EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARAGPLYTIFGVVIIPDNWFDPWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:14; variable domain underlined; CDRs double underlined).

[0138] Heavy Chain Variable Domain EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARAGPLYTIFGVVIIPDNWFDPWGQGTLVTVSS (Sequence number: 68).

[0139] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO: 14 or 68, where residue 1 of SEQ ID NO: 14 or 68 is Q instead of E.

[0140] 2C1 Heavy Chain CDR CDR-H1: ELSMH (SEQ ID NO: 34) CDR-H2: GFDPEDGETIYAQKFQG (SEQ ID NO: 35) CDR-H3: AGPLYTIFGVVIIPDNWFDP (sequence number: 36).

[0141] Antibody 2C1 light chain (Q1E) (lambda) Light chain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSGVVFGGGTQLIIL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:15; variable domains underlined; CDRs double underlined).

[0142] Light Chain Variable Domain ESVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSGVVFGGGTQLIIL (Sequence number: 76).

[0143] The formulations disclosed herein can be used with antibodies and antigen-binding fragments thereof comprising SEQ ID NO: 15 or 76, where residue 1 of SEQ ID NO: 15 or 76 is Q instead of E.

[0144] 2C1 light chain CDRs CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 37) CDR-L2: GNSNRPS (SEQ ID NO: 38) CDR-L3: QSYDSSLSGSGVV (sequence number: 39).

[0145] 2C1 heavy and light chain CDRs or 2C1 V H and V L Alternatively, antibodies and antigen-binding fragments thereof comprising 2C1 heavy and light chains (or variants thereof, such as those described herein) may be referred to as "2C1."

[0146] In various embodiments of the antibody or antigen-binding fragment thereof, the C-terminal lysine of the heavy chain immunoglobulin is absent.

[0147] Thus, in some embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain immunoglobulin, where the light chain immunoglobulin comprises the amino acid sequence set forth in SEQ ID NO:3, 4, 5, 6, 7, 11, 13, 15 or 45; and / or the heavy chain immunoglobulin comprises the amino acid sequence set forth in SEQ ID NO:1, 2, 8, 9, 10, 12, 14, 44, 79, 80, 81, 82, 83, 84, 85 or 86.

[0148] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:1 or 79; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:3.

[0149] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:4.

[0150] In another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:5.

[0151] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:6.

[0152] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:7.

[0153] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:2 or 80; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:3.

[0154] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:8 or 82; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:11.

[0155] In another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:9 or 83; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:11.

[0156] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 10 or 84; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 11.

[0157] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 12 or 85; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 13.

[0158] In further embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 14 or 86; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 15.

[0159] In certain further embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain, wherein the light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 70, 71, 72, 73, 58, 74, 75, 76, or 77, and / or the heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 63, 57, 64, 65, 66, 67, 68, or 69.

[0160] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:63; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:70.

[0161] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:71.

[0162] In another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:72.

[0163] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:73.

[0164] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0165] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:70.

[0166] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:64; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:74.

[0167] In another embodiment, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:65; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:74.

[0168] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:66; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:74.

[0169] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:67; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:75.

[0170] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:68; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:76.

[0171] In a further embodiment, the antibody or antigen-binding fragment thereof that binds to ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 1E1 (e.g., SEQ ID NOs: 19-21). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising CDR-H1, CDR-H2 and CDR-H3 of 1E1 (e.g., SEQ ID NO:16-18). H ) domain;

[0172] In a further embodiment, the antibody or antigen-binding fragment thereof that binds to ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 2A6 (e.g., SEQ ID NOs:25-27). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising the CDR-H1, CDR-H2 and CDR-H3 of 2A6 (e.g., SEQ ID NO:22-24). H ) domain;

[0173] In a further embodiment, the antibody or antigen-binding fragment thereof that binds to ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 3G7 (e.g., SEQ ID NOs: 31-33). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising the CDR-H1, CDR-H2 and CDR-H3 of 3G7 (e.g., SEQ ID NO:28-30). H ) domain;

[0174] In a further embodiment, the antibody or antigen-binding fragment thereof that binds to ILT4 has an immunoglobulin light chain variable (VV) domain comprising CDR-L1, CDR-L2 and CDR-L3 of 2C1 (e.g., SEQ ID NOs: 37-39). L ) domain; and an immunoglobulin heavy chain variable (V) domain comprising CDR-H1, CDR-H2 and CDR-H3 of 2C1 (e.g., SEQ ID NOs: 34-36). H ) domain;

[0175] In one embodiment, the antibody or antigen-binding fragment comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNSNRPS (SEQ ID NO: 49), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0176] In another embodiment, the antibody or antigen-binding fragment comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQSNRPS (SEQ ID NO: 50), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0177] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GESNRPS (SEQ ID NO: 51), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0178] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). Hdomain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0179] In one embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNANRPS (SEQ ID NO: 53), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0180] In another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQANRPS (SEQ ID NO: 54), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0181] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GEANRPS (SEQ ID NO: 55), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0182] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDANRPS (SEQ ID NO: 56), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0183] In one embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNSNRPS (SEQ ID NO: 49), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0184] In another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQSNRPS (SEQ ID NO: 50), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0185] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GESNRPS (SEQ ID NO: 51), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0186] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0187] In one embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GNANRPS (SEQ ID NO: 53), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0188] In another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). Hdomain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GQANRPS (SEQ ID NO: 54), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0189] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GEANRPS (SEQ ID NO: 55), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0190] In yet another embodiment, the antibody or antigen-binding fragment thereof comprises CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18). H domain; and / or CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDANRPS (SEQ ID NO: 56), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21) L domain;

[0191] The formulations disclosed herein bind to ILT4 and have the V L Domains (e.g., SEQ ID NOs: 70, 71, 72, 73, 58, or 77) and / or V of antibody 1E1 H It may be used in conjunction with an antibody or antigen-binding fragment thereof that contains the domain (e.g., SEQ ID NO: 63, 57, or 69).

[0192] The formulations disclosed herein bind to ILT4 and are LDomain (e.g., SEQ ID NO:74) and / or V of antibody 2A6 H It may be used in conjunction with an antibody or antigen-binding fragment thereof comprising the domain (e.g., SEQ ID NO: 64, 65 or 66).

[0193] The formulations disclosed herein bind to ILT4 and are L Domains (e.g., SEQ ID NO:75) and / or V of antibody 3G7 H The antibody or antigen-binding fragment thereof may be used in combination with an antibody comprising the domain (eg, SEQ ID NO:67).

[0194] The formulations disclosed herein bind to ILT4 and have the V L Domain (e.g., SEQ ID NO:76) and / or V of antibody 2C1 H It may be used in conjunction with an antibody or antigen-binding fragment thereof that contains the domain (eg, SEQ ID NO:68).

[0195] The formulations disclosed herein may be used with antibodies or antigen-binding fragments thereof that bind ILT4 and include the light chain immunoglobulin of antibody 1E1 (e.g., SEQ ID NO:3, 4, 5, 6, 7 or 45) and / or the heavy chain immunoglobulin of antibody 1E1 (e.g., SEQ ID NO:1, 2, 44, 79, 80 or 81).

[0196] The formulations disclosed herein may be used with antibodies or antigen-binding fragments thereof that bind ILT4 and include the light chain immunoglobulin of antibody 2A6 (e.g., SEQ ID NO:11) and / or the heavy chain immunoglobulin of antibody 2A6 (e.g., SEQ ID NOs:8, 9, 10, 82, 83 or 84).

[0197] The formulations disclosed herein may be used with an antibody or antigen-binding fragment thereof that binds ILT4 and includes the light chain immunoglobulin of antibody 3G7 (e.g., SEQ ID NO:13) and / or the heavy chain immunoglobulin of antibody 3G7 (e.g., SEQ ID NO:12 or 85).

[0198] The formulations disclosed herein may be used with an antibody or antigen-binding fragment thereof that binds ILT4 and comprises the light chain immunoglobulin of antibody 2C1 (e.g., SEQ ID NO:15) and / or the heavy chain immunoglobulin of antibody 2C1 (e.g., SEQ ID NO:14 or 86).

[0199] The formulations disclosed herein can be used with antibodies that consist of two heavy chains and two light chains, where each light chain is a V of antibody 1E1, 2A6, 3G7, or 2C1. L or light chain immunoglobulin, each heavy chain comprising the V of antibody 1E1, 2A6, 3G7 or 2C1 H or heavy chain immunoglobulins.

[0200] In one embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57.

[0201] In another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58, each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57, and the light chains further comprise the amino acid sequence set forth in SEQ ID NO:90.

[0202] In yet another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57, wherein the heavy chains further comprise the amino acid sequence set forth in SEQ ID NO:89.

[0203] In yet another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:58 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:57, wherein the light chain further comprises the amino acid sequence set forth in SEQ ID NO:90 and the heavy chain further comprises the amino acid sequence set forth in SEQ ID NO:89.

[0204] In one embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain comprises the amino acid sequence set forth in SEQ ID NO:7 and each heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2.

[0205] In another embodiment, the antibody consists of two heavy chains and two light chains, wherein each light chain consists of the amino acid sequence set forth in SEQ ID NO:7 and each heavy chain consists of the amino acid sequence set forth in SEQ ID NO:2.

[0206] In one embodiment, the antibody or antigen-binding fragment thereof comprises L (with or without a signal sequence), e.g., one, two, three, four, five, six, seven, eight, nine, ten or more conservative or non-conservative amino acid substitutions, such as V in any of SEQ ID NOs: 58 or 70-77 L and / or V H 57 or 63-69 (with or without a signal sequence), e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more conservative or non-conservative amino acid substitutions and still bind to ILT4 H ;includes.

[0207] The formulations disclosed herein can be used with polypeptides that include an amino acid sequence disclosed herein (e.g., SEQ ID NOs:1-39, 44, 45, 47-58, 63-77 or 79-86), as well as with polypeptides that include said amino acid sequences having up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20 or more conservative or non-conservative amino acid substitutions therein.

[0208] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain immunoglobulin, wherein the light chain immunoglobulin has at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:3, 4, 5, 6, 7, 11, 13, 15 or 45, and / or the heavy chain immunoglobulin has at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:1, 2, 8, 9, 10, 12, 14, 44, 79, 80, 81, 82, 83, 84, 85 or 86.

[0209] In some embodiments, the antibody or antigen-binding fragment thereof comprises a light chain immunoglobulin, a heavy chain immunoglobulin, or both a light chain immunoglobulin and a heavy chain immunoglobulin, wherein the light chain immunoglobulin comprises a light chain variable domain having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 70, 71, 72, 73, 58, 74, 75, 76 or 77, and / or the heavy chain immunoglobulin comprises a heavy chain variable domain having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 63, 57, 64, 65, 66, 67, 68 or 69.

[0210] In one embodiment, the immunoglobulin heavy chain of the anti-ILT4 antibody or antigen-binding fragment of the invention is operably linked to a signal sequence (e.g., a signal sequence comprising the amino acid sequence MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 41)) and / or the immunoglobulin light chain of the anti-ILT4 antibody or antigen-binding fragment of the invention is operably linked to a signal sequence (e.g., a signal sequence comprising the amino acid sequence MSVPTQVLGLLLLWLTDARC (SEQ ID NO: 42)).

[0211] In one embodiment, the N-terminal glutamine (Q) of the immunoglobulin chains (e.g., heavy and / or light chains) described herein is replaced with pyroglutamic acid. In one embodiment, the N-terminal Q of the heavy chain immunoglobulin is replaced with pyroglutamic acid. In another embodiment, the N-terminal Q of the light chain immunoglobulin is replaced with pyroglutamic acid. In yet another embodiment, the N-terminal Q of the heavy chain immunoglobulin and the N-terminal Q of the heavy chain immunoglobulin are replaced with pyroglutamic acid.

[0212] The formulations disclosed herein can also be used with any of the anti-ILT4 antibodies or antigen-binding fragments thereof disclosed herein (e.g., 1E1, 2A6, 3G7, or 2C1) and an antibody or antigen-binding fragment that binds to the same epitope of ILT4 (e.g., human ILT4). In one embodiment, the epitope is LYREKKSASW (SEQ ID NO:59). In another embodiment, the epitope is TRIRPEL (SEQ ID NO:60). In yet another embodiment, the epitope is NGQF (SEQ ID NO:61). In yet another embodiment, the epitope is HTGRYGCQ (SEQ ID NO:62). In certain embodiments, the antibody or antigen-binding fragment thereof binds to the same epitope of human ILT4 as an antibody or antigen-binding fragment thereof comprising the heavy chain and light chain amino acid sequences set forth in SEQ ID NOs:1 and 3; 2 and 4; 2 and 5; 2 and 6; 2 and 7; 2 and 3; 8 and 11; 9 and 11; 10 and 11; 12 and 13; 14 and 15; 79 and 3; 80 and 4; 80 and 5; 80 and 6; 80 and 7; 80 and 3; 82 and 11; 83 and 11; 84 and 11; 85 and 13; and 86 and 15; respectively. In some embodiments, the antibody or antigen-binding fragment thereof binds to the same epitope of human ILT4 as an antibody or antigen-binding fragment thereof comprising the heavy chain variable domain amino acid sequences and the light chain variable domain amino acid sequences set forth in SEQ ID NOs: 63 and 70; 57 and 71; 57 and 72; 57 and 73; 57 and 58; 57 and 70; 64 and 74; 65 and 74; 66 and 74; 67 and 75; 68 and 76; respectively.

[0213] The formulations disclosed herein can be used with antibodies or antigen-binding fragments that cross-block the binding of any of the anti-ILT4 antibodies or antigen-binding fragments thereof disclosed herein (e.g., 1E1, 2A6, 3G7 or 2C1) to ILT4 (e.g., human ILT4) or with antibodies and antigen-binding fragments that compete with the binding of any of the anti-ILT4 antibodies or antigen-binding fragments thereof disclosed herein (e.g., 1E1, 2A6, 3G7 or 2C1) to ILT4 (e.g., human ILT4). The cross-blocking antibodies and antigen-binding fragments thereof discussed herein can be confirmed based on their ability to block any of the antibodies or fragments specifically described herein from binding to ILT4 in binding assays (e.g., biolayer interferometry (BLI; e.g., FORTEBIO OCTET binding assay; Pall ForteBio Corp; Menlo Park, CA), surface plasmon resonance (SPR), BIACore, ELISA, flow cytometry). For example, in an embodiment of the invention, when using BLI, the tip of a fiber optic probe is coated with a ligand (e.g., ILT4) and functions as a biosensor, where binding of an anti-ILT4 antibody or antigen binding fragment to ILT4 changes the interference pattern of white light reflected from the ILT4-bound probe layer and the internal reference layer. This shift is indicative of ILT4 / anti-ILT4 binding. In an embodiment of the invention, the ILT4-coated tip is immersed in a solution of analyte containing antibodies or antigen binding fragments, for example in the wells of either a 96-well plate or a 384-well plate. In an embodiment of the invention, the plate is shaken during reading to create orbital flow. To read the assay, white light is directed down the length of the fiber. As described above, interference of the light reflected from the reference layer with the immobilization surface containing ILT4 on the tip produces a characteristic pattern of light reflected back up the fiber. When molecules bind to the immobilized sensor surface, the pattern changes in proportion to the extent of binding.For example, an assay can be used in which ILT4 (e.g., human ILT4) protein is immobilized on a BLI probe or plate, a reference anti-ILT4 antibody or fragment is bound to ILT4 (e.g., at saturating concentration), and a test anti-ILT4 antibody or fragment is added. The ability of the test antibody to compete with the reference antibody for ILT4 binding is then measured. In a BLI format, the light interference of the ILT4 complex is monitored to confirm whether the test antibody effectively competes with the reference antibody. For example, nanometers of the wavelength shift of light over time are monitored, where the shift indicates additional binding and lack of cross-blocking of the test antibody. In an embodiment of the invention, in a BLI format, if no additional binding of the test antibody is observed, cross-blocking between the antibodies is deemed to have occurred qualitatively. In an embodiment of the invention, as a control, cross-blocking of the reference antibody with itself is confirmed; where if the reference antibody is able to cross-block itself from ILT4 binding, the assay is confirmed to be working correctly. The ability of the test antibody to inhibit the binding of anti-ILT4 antibody or fragment 1E1, 2A6, 3G7 or 2C1 to ILT4 (e.g. human ILT4) indicates that the test antibody is capable of cross-blocking the antibody or fragment for binding to ILT4 (e.g. human ILT4) and therefore, in some cases, may bind to the same epitope on ILT4 (e.g. human ILT4) as 1E1, 2A6, 3G7 and / or 2C1. As mentioned above, antibodies and fragments that bind to the same epitope as any of the anti-ILT4 antibodies or fragments of the invention also form part of the invention. In one embodiment of the invention, BLI is performed in a sandwich format, where a reference anti-ILT4 antibody or antigen-binding fragment is immobilized on a probe and then binds to ILT4. The test anti-ILT4 antibody or antigen-binding fragment is then tested for its ability to block the binding of the reference antibody or fragment.

[0214] In certain embodiments, the antibody or antigen-binding fragment thereof competes for binding to human ILT4 with an antibody or fragment comprising the heavy chain and light chain amino acid sequences set forth in SEQ ID NOs:1 and 3; 2 and 4; 2 and 5; 2 and 6; 2 and 7; 2 and 3; 8 and 11; 9 and 11; 10 and 11; 12 and 13; 14 and 15; 79 and 3; 80 and 4; 80 and 5; 80 and 6; 80 and 7; 80 and 3; 82 and 11; 83 and 11; 84 and 11; 85 and 13; and 86 and 15; respectively. In some embodiments, the antibody or antigen-binding fragment thereof competes for binding to human ILT4 with an antibody or fragment comprising the heavy chain variable domain amino acid sequences and the light chain variable domain amino acid sequences set forth in SEQ ID NOs: 63 and 70; 57 and 71; 57 and 72; 57 and 73; 57 and 58; 57 and 70; 64 and 74; 65 and 74; 66 and 74; 67 and 75; 68 and 76; respectively.

[0215] The formulations disclosed herein can be used with anti-ILT4 antibodies and antigen-binding fragments thereof that include N-linked glycans typically added to immunoglobulins produced in Chinese hamster ovary cells (CHO N-linked glycans) or with anti-ILT4 antibodies and antigen-binding fragments thereof that include N-linked glycans typically added to immunoglobulins produced in artificial yeast cells (e.g., Pichia pastoris) (artificial yeast N-linked glycans). For example, in embodiments, the anti-ILT4 antibodies and antigen-binding fragments thereof include one or more of "artificial yeast N-linked glycans" or "CHO ​​N-linked glycans" (e.g., G0 and / or G0-F and / or G1 and / or G1-F and / or G2-F and / or Man5). In embodiments, the anti-ILT4 antibodies and antigen-binding fragments thereof include artificial yeast N-linked glycans (i.e., G0 and / or G1 and / or G2, optionally further including Man5). In embodiments, the anti-ILT4 antibodies and antigen-binding fragments thereof comprise CHO N-linked glycans (i.e., G0-F, G1-F and G2-F, optionally further including G0 and / or G1 and / or G2 and / or Man5). In embodiments, about 80% to about 95% (e.g., about 80-90%, about 85%, about 90% or about 95%) of the total N-linked glycans on the anti-ILT4 antibodies and antigen-binding fragments thereof are artificial yeast N-linked glycans or CHO N-linked glycans. See, Nett et al. Yeast. 28(3): 237-252 (2011); Hamilton et al. Science. 313(5792): 1441-1443 (2006); Hamilton et al. Curr Opin Biotechnol. 18(5): 387-392 (2007). For example, in embodiments, the artificial yeast cell is GFI5.0 or YGLY8316, or a strain described in U.S. Patent No. 7,795,002 or Zha et al., Methods Mol Biol. 988:31-43 (2013). See also International Patent Application Publication No. WO2013 / 066765.

[0216] Anti-ILT4 antibodies and antigen-binding fragments thereof (e.g., 1E1, 2A6, 3G7, and / or 2C1) that can be used with the various formulations disclosed herein can also be engineered to include modifications within the Fc region, typically to alter one or more functional properties of the antibody (e.g., serum half-life, complement fixation, Fc receptor binding, and / or effector function (e.g., antigen-dependent cellular cytotoxicity), etc.). Additionally, antibodies and antigen-binding fragments thereof (e.g., 1E1, 2A6, 3G7, and / or 2C1) that can be used with the various formulations disclosed herein can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody, such as a labeled antibody or antibody-drug conjugate) or modified to alter glycosylation, also to alter one or more functional properties of the antibody.

[0217] In some embodiments of the various formulations disclosed herein, the antibody or antigen-binding fragment is a monoclonal antibody.

[0218] Pharmaceutical preparations The pharmaceutical formulations described herein may retain the physical, chemical and / or biological stability of the anti-ILT4 antibody (e.g., 1E1, 2A6, 3G7 or 2C1) or antigen-binding fragment thereof during storage (e.g., at least 6, 12, 24 or 36 months at about 5°C, at least 6, 12, 24 or 36 months at -20°C, or at least 6, 12, 24 or 36 months at -70°C) and / or under various stress conditions (e.g., agitation, freeze-thaw cycles, or light exposure).

[0219] In one aspect, provided herein are various formulations of an anti-ILT4 monoclonal antibody (e.g., 1E1, 2A6, 3G7, or 2C1) or an antigen-binding fragment thereof, comprising: (i) an anti-ILT4 antibody or antigen-binding fragment thereof (e.g., 1E1, 2A6, 3G7, or 2C1); (ii) a buffer (e.g., an L-histidine buffer or an acetate buffer); (iii) a non-reducing sugar (e.g., sucrose); (iv) a non-ionic detergent (e.g., PS-80); and (v) an antioxidant (e.g., L-methionine).

[0220] Buffering agents that can be used in the pharmaceutical formulations disclosed herein include, but are not limited to, succinate (sodium or potassium), L-histidine, phosphate (sodium or potassium), Tris (tris(hydroxymethyl)aminomethane), diethanolamine, citrate (sodium), acetate (sodium), etc. In some embodiments of the formulations, the buffering agent is present in the formulation at a concentration of about 1-20 mM (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mM).

[0221] The buffers of the various formulations described herein have a pH range of about 4.5 to about 7.0, about 4.5 to about 6.8, about 5.0 to about 6.8, about 5.0 to about 6.5, about 5.0 to about 6.0, and about 5.5 to about 6.0. In arriving at a representative formulation, L-histidine and acetate buffers in the pH range of 5.0 to 6.8 were examined for suitability. When a range of pH values ​​is described, such as "a pH of about pH 5.5 to about 6.0", the range is intended to include the described value. For example, the range of about 5.0 to about 6.0 includes 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, and 6.0. When the anti-ILT4 formulation described herein is a lyophilized formulation, the pH refers to the pH after reconstitution of the lyophilized formulation. pH is typically measured with a standard glass bulb pH meter at 25° C. As used herein, a solution containing "a histidine buffer of pH X" refers to a solution containing a histidine buffer at pH X, i.e., the pH is intended to refer to the pH of the solution.

[0222] In some embodiments, the anti-ILT4 formulation comprises a non-reducing sugar. As used herein, a "non-reducing sugar" is a sugar that does not contain or cannot be converted to contain a free aldehyde or ketone group and therefore cannot act as a reducing agent. Examples of non-reducing sugars include, but are not limited to, disaccharides such as sucrose and trehalose. In an embodiment of the invention, the non-reducing sugar is present in an amount of about 1% (w / v) to about 10% (w / v) (about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10% (w / v)). In another embodiment, the non-reducing sugar is present in an amount of about 6% (w / v) to about 8% (w / v) (about 6, 7, or 8% (w / v)). In a further embodiment, the non-reducing sugar is present in an amount of about 6% (w / v). In a further embodiment, the non-reducing sugar is present in an amount of about 7% (w / v). In a further embodiment, the non-reducing sugar is present in an amount of about 8% (w / v). In one embodiment, the non-reducing sugar is sucrose, trehalose or raffinose. In a further embodiment, sucrose is present in an amount of about 6% (w / v) to about 8% (w / v). In one embodiment, sucrose is present at about 6% (w / v). In one embodiment, sucrose is present at about 7% (w / v). In one embodiment, sucrose is present at about 8% (w / v).

[0223] In certain embodiments, the anti-ILT4 formulation further comprises a surfactant. As used herein, a "surfactant" is a surface active agent that is amphiphilic in nature. Surfactants can be added to the formulations herein to provide stability, reduce and / or prevent aggregation, or prevent and / or inhibit protein damage under process conditions such as purification, filtration, lyophilization, transportation, storage and delivery. In some embodiments, surfactants can be useful to provide additional stability to the active ingredient(s).

[0224] Nonionic surfactants that can be used in the formulations of the invention include, but are not limited to, polyoxyethylene sorbitan fatty acid esters (polysorbates, sold under the trade name Tween® (Uniquema Americas LLC, Wilmington, DE)), such as polysorbate-20 (PS-20, polyoxyethylene sorbitan monolaurate), polysorbate-40 (PS-40, polyoxyethylene sorbitan monopalmitate), polysorbate-60 (PS-60, polyoxyethylene sorbitan monostearate), and polysorbate-80 (PS-80, polyoxyethylene sorbitan monooleate); polyoxyethylene alkyl ethers, such as Brij® 58 (Uniquema Americas LLC, Wilmington, DE) and Brij® 35; poloxamers (e.g., poloxamer 188); Triton® X-100 (Union Carbide Corp., Houston, TX) and Triton® X-114; NP40; Span 20, Span 40, Span 60, Span 65, Span 80 and Span 85; copolymers of ethylene and propylene glycol (e.g., the pluronic® series of nonionic surfactants, such as pluronic® F68, pluronic® 10R5, pluronic® F108, pluronic® F127, pluronic® F38, pluronic® L44, pluronic® L62 (BASF Corp., Ludwigshafen, Germany); and sodium dodecyl sulfate (SDS). In one embodiment, the nonionic surfactant is PS-80 or PS-20. In one embodiment, the nonionic surfactant is PS-20. In another embodiment, the nonionic surfactant is PS-80.

[0225] The amount of non-ionic surfactant included in the formulation is sufficient to perform the desired function, i.e., the minimum amount required to stabilize the active pharmaceutical ingredient (i.e., anti-ILT4 antibody or antigen-binding fragment thereof (e.g., 1E1, 2A6, 3G7, or 2C1)) in the formulation. All percentages of the non-ionic surfactant are listed in % (w / v). Typically, the surfactant is present at a concentration of about 0.008% to about 0.1% (w / v). In some embodiments of this aspect of the invention, the surfactant is present in the formulation in an amount of about 0.01% to about 0.1%, about 0.01% to about 0.09%, about 0.01% to about 0.08%, about 0.01% to about 0.07%, about 0.01% to about 0.06%, about 0.01% to about 0.05%, about 0.01% to about 0.04%, about 0.01% to about 0.03%, about 0.01% to about 0.02%, about 0.015% to about 0.04%, about 0.015% to about 0.03%, about 0.015% to about 0.02%, about 0.02% to about 0.04%, about 0.02% to about 0.035%, or about 0.02% to about 0.03%. In alternative embodiments, the surfactant is present in an amount of about 0.01%, about 0.015%, about 0.02%, about 0.025%, about 0.03%, about 0.035% or about 0.04%.

[0226] In certain embodiments, the formulations described herein comprise about 0.01% to about 0.04% (w / v) PS-80. In one embodiment, the formulations described herein comprise PS-80 in an amount of about 0.01%. In one embodiment, the amount of PS-80 is about 0.015%. In another embodiment, the amount of PS-80 is about 0.02%. In a further embodiment, the amount of PS-80 is about 0.025%. In another embodiment, the amount of PS-80 is about 0.03%. In a further embodiment, the amount of PS-80 is about 0.035%. In another embodiment, the amount of PS-80 is about 0.04%. In a further embodiment, the amount of PS-80 is about 0.045%.

[0227] The formulations described herein further comprise methionine or a pharma- ceutically acceptable salt thereof as an antioxidant. In one embodiment, the methionine is L-methionine. In another embodiment, the methionine is a pharma- ceutically acceptable salt of L-methionine, e.g., methionine HCl. In one embodiment of the invention, methionine is present in the formulation at a concentration of about 1 to 20 mM (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 mM). In another embodiment, methionine is present at about 5 mM to about 10 mM (5, 6, 7, 8, 9, and 10 mM). In another embodiment, methionine is present at about 10 mM.

[0228] Accordingly, in one aspect, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM of a buffer; (iii) about 6% to about 8% weight / volume (w / v) of a non-reducing sugar; (iv) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (v) about 1 mM to about 20 mM of an antioxidant.

[0229] In certain embodiments, the buffer is selected from the group consisting of an L-histidine buffer, an acetate buffer, and a citrate buffer. In one embodiment, the buffer is an L-histidine buffer. In another embodiment, the buffer is an acetate buffer. In yet another embodiment, the buffer is a citrate buffer.

[0230] In some embodiments, the non-reducing sugar is sucrose.

[0231] In certain embodiments, the non-ionic surfactant is PS-80 or PS-20. In one embodiment, the non-ionic surfactant is PS-80. In another embodiment, the non-ionic surfactant is PS-20.

[0232] In some embodiments, the antioxidant is L-methionine.

[0233] Accordingly, in another aspect, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM L-histidine buffer; (iii) about 6% to about 8% (w / v) sucrose; (iv) about 0.01% to about 0.10% (w / v) PS-80; and (v) about 1 mM to about 20 mM L-methionine.

[0234] In some embodiments, the formulation comprises about 8 mM to about 12 mM L-histidine buffer.

[0235] In certain embodiments, the formulation comprises about 5 mM to about 10 mM L-methionine.

[0236] In another embodiment, the formulation comprises about 0.01% to about 0.05% (w / v) PS-80.

[0237] In yet another embodiment, the formulation comprises about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 15 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL, or about 150 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 12.5 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 15 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 25 mg / mL. In one embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 50 mg / mL. In another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 75 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 100 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 125 mg / mL. In yet another embodiment, the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 150 mg / mL.

[0238] Thus, in one particular embodiment, the formulation comprises about 25 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0239] In another specific embodiment, the formulation comprises about 50 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0240] In yet another specific embodiment, the formulation comprises about 75 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0241] In yet another specific embodiment, the formulation comprises about 100 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0242] In yet another specific embodiment, the formulation comprises about 125 mg / mL of anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80 and about 10 mM L-methionine.

[0243] In certain embodiments of the various formulations provided herein, the formulation has a pH of about 5.0 to about 6.8. In some embodiments, the formulation has a pH of about 5.5 to about 6.0. In other embodiments, the formulation has a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.0. In one embodiment, the formulation has a pH of about 5.5. In another embodiment, the formulation has a pH of about 5.6. In yet another embodiment, the formulation has a pH of about 5.7. In yet another embodiment, the formulation has a pH of about 5.8. In another embodiment, the formulation has a pH of about 5.9. In yet another embodiment, the formulation has a pH of about 6.0.

[0244] Thus, in one particular embodiment, the pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprises: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer, pH about 5.5; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) polysorbate 80; and (v) about 10 mM L-methionine.

[0245] In certain embodiments of the various formulations disclosed herein, the anti-ILT4 antibody or antigen-binding fragment thereof is an anti-ILT4 antibody or antigen-binding fragment thereof described herein (e.g., 1E1, 2A6, 3G7, or 2C1). In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 1E1 or a variant thereof. In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 2A6 or a variant thereof. In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 3G7 or a variant thereof. In some embodiments of the formulation, the anti-ILT4 antibody or antigen-binding fragment thereof is 2C1 or a variant thereof.

[0246] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0247] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0248] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0249] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0250] In yet another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80, and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0251] In certain embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57, and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58.

[0252] In some embodiments of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0253] In another embodiment of the various formulations provided herein, the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0254] Accordingly, in some embodiments, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the CDR-H1:GYYWS (sequence and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0255] In certain embodiments, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof is The original binding fragment comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0256] In another embodiment, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57 and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0257] In yet another embodiment, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0258] In yet another embodiment, provided herein is a pharmaceutical formulation for an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0259] In some embodiments, provided herein is a pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:57 and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:58.

[0260] In some embodiments, provided herein is a pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0261] In some embodiments, provided herein is a pharmaceutical formulation of an anti-ILT4 antibody or antigen-binding fragment thereof comprising: (i) about 25 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer; (iii) about 7% (w / v) sucrose; (iv) about 0.025% (w / v) PS-80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:80 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0262] In one embodiment, the formulation is a liquid formulation. In another embodiment, the liquid formulation is stored at about 3° C. to about 5° C. In yet another embodiment, the liquid formulation is frozen at at least −70° C. or below. In yet another embodiment, the liquid formulation is a reconstituted solution from a lyophilized formulation.

[0263] In certain embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, (i) the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) the OD 0.350-500(iii) the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and / or the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% as measured by high performance ion exchange chromatography; (iv) the number of subvisible particles of particles at least 2 μm in size is at most about 3500 as measured by microflow imaging; and / or (v) the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping.

[0264] In some embodiments, the % monomer of the anti-ILT4 antibody, or antigen-binding fragment thereof, is at least about 99% when measured by ultra performance size exclusion chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months.

[0265] In some embodiments, the formulation is stored at about 3° C. to about 5° C. for 6 months and then the OD 350-500 When measured at 100° C., the turbidity of the formulation is at most about 0.135.

[0266] In some embodiments, the % major peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography after storage of the formulation at about 3° C. to about 5° C. for 6 months. In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the % major peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14% when measured by high performance ion exchange chromatography.

[0267] In some embodiments, after storage of the formulation at about 3° C. to about 5° C. for 6 months, the number of sub-visible particles having a size of at least 2 μm is up to about 3500 as measured by microflow imaging.

[0268] In some embodiments, the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, the % oxidation of one amino acid residue selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping after storage of the formulation at about 3° C. to about 5° C. for 6 months. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of two amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of three amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of four amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%. In one embodiment, when the formulation is stored at about 3° C. to about 5° C. for 6 months and then measured by reduced peptide mapping, the % oxidation rate of all five amino acid residues in the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody described in SEQ ID NO: 2 or 80 is less than about 4%.

[0269] The formulations described herein can be produced as liquid formulations. Liquid antibody formulations can be produced by buffer exchange of the drug substance (e.g., anti-ILT4 monoclonal antibody) in liquid form (e.g., anti-ILT4 monoclonal antibody in an aqueous pharmaceutical formulation) into a desired buffer as the final step in the purification process. In this embodiment, there is no lyophilization step. The drug substance in the final buffer is concentrated to the desired concentration. Excipients such as sucrose and PS80 are added to the drug substance and diluted with an appropriate buffer to the final protein concentration. The final formulated drug substance is filtered using a 0.22 μm filter and filled into the final container (e.g., glass vial).

[0270] The formulations described herein can also be reconstituted from lyophilized formulations. Lyophilized formulations of therapeutic proteins offer several advantages. Lyophilized formulations generally offer better chemical stability than solution formulations, and therefore have increased half-life. Lyophilized formulations can also be reconstituted at different concentrations depending on clinical factors such as the route of administration or dosing. For example, lyophilized formulations are reconstituted at higher concentrations (i.e., smaller volumes) if subcutaneous administration is required, or at lower concentrations if administered intravenously. Higher concentrations may be necessary if a higher dose is required for a particular subject, particularly for subcutaneous administration where the injection volume must be minimized. One such lyophilized antibody formulation is disclosed in U.S. Pat. No. 6,267,958, which is incorporated herein by reference in its entirety. Another lyophilized formulation of a therapeutic protein is disclosed in U.S. Pat. No. 7,247,707, which is incorporated herein by reference in its entirety.

[0271] Typically, lyophilized formulations are prepared with the intention of reconstituting a high concentration of drug product (DP, in an exemplary embodiment, an anti-ILT4 antibody or antigen-binding fragment thereof), i.e., with a low volume of water. Subsequent dilution with water or an isotonic buffer can then be easily used to reduce the DP to a low concentration. Typically, excipients are included in the lyophilized formulations of the invention at levels that result in a near-isotonic formulation when reconstituted at a high DP concentration, e.g., for subcutaneous administration. Reconstitution with a large amount of water to reduce the DP concentration necessarily reduces the tonicity of the reconstituted solution, but such reduction may be of little importance for non-subcutaneous administration (e.g., intravenous administration). If isotonicity is desired at a lower DP concentration, the lyophilized powder can be reconstituted with a standard low volume of water and then diluted with an isotonic diluent, such as 0.9% sodium chloride.

[0272] The lyophilized formulations of the present invention are formed by lyophilizing the pre-lyophilization solution. Lyophilization is accomplished by freezing the formulation followed by sublimation of water at a temperature suitable for primary drying. Under these conditions, the product temperature is below the eutectic or collapse temperature of the formulation. Typically, the shelf temperature for primary drying ranges from about -30 to 25°C at an appropriate pressure, typically in the range of about 50 to 250 mTorr, provided that the product remains frozen during primary drying. The formulation, the size and type of container (e.g., glass vial) in which the sample is placed, and the amount of liquid determine the time required for drying, which can range from a few hours to several days (e.g., 40 to 60 hours). The secondary drying step can be carried out at about 0 to 40°C, depending primarily on the type and size of the container and the type of protein used. The secondary drying time is determined by the desired residual moisture level in the product and typically takes at least about 5 hours. Typically, the moisture content of the lyophilized formulation is less than about 5%, preferably less than about 3%. The pressure can be the same as that employed in the primary drying step. The lyophilization conditions can vary depending on the formulation and vial size.

[0273] In some cases, to avoid transfer steps, it may be desirable to lyophilize the protein formulation in the container in which the protein reconstitution will be performed, e.g., a 3, 5, 10, 20, 50, or 100 cc vial.

[0274] The lyophilized formulation is reconstituted prior to administration. The protein may be reconstituted at a concentration of about 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, or 100 mg / mL, or at a higher concentration up to about 500 mg / mL, for example, 150 mg / mL, 200 mg / mL, 250 mg / mL, or 300 mg / mL. In one embodiment, the protein concentration after reconstitution is about 10-300 mg / mL. In one embodiment, the protein concentration after reconstitution is about 20-250 mg / mL. In one embodiment, the protein concentration after reconstitution is about 150-250 mg / mL. In one embodiment, the protein concentration after reconstitution is about 180-220 mg / mL. In one embodiment, the protein concentration after reconstitution is about 50-150 mg / mL. In one embodiment, the protein concentration after reconstitution is about 150 mg / mL. In one embodiment, the protein concentration after reconstitution is about 125 mg / mL. In one embodiment, the protein concentration after reconstitution is about 100 mg / mL. In one embodiment, the protein concentration after reconstitution is about 75 mg / mL. In one embodiment, the protein concentration after reconstitution is about 50 mg / mL. In one embodiment, the protein concentration after reconstitution is about 25 mg / mL. High protein concentrations are particularly useful when subcutaneous delivery of the reconstituted formulation is intended. However, for other routes of administration, such as intravenous administration, lower concentrations of protein may be desired (e.g., about 5-50 mg / mL).

[0275] Reconstitution is usually carried out at a temperature of about 25° C. to ensure complete hydration, although other temperatures may be used if desired. The time required for reconstitution depends, for example, on the type of diluent, the amount of excipients, and the protein. Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffered solution (e.g., phosphate buffered saline), sterile saline, Ringer's solution, or dextrose solution.

[0276] How to use In another aspect, provided herein is a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of any formulation described herein. In some specific embodiments of this method, the formulation is administered to the subject via intravenous administration. In another embodiment, the formulation is administered to the subject by subcutaneous administration. In one embodiment, the invention encompasses a method for treating cancer in a human patient, comprising administering to the patient any formulation described herein.

[0277] In any of the methods of the invention, the cancer may be selected from the group consisting of melanoma, lung cancer, head and neck cancer, bladder cancer, breast cancer, gastrointestinal cancer, multiple myeloma, hepatocellular carcinoma, lymphoma, renal cancer, mesothelioma, ovarian cancer, esophageal cancer, anal cancer, biliary tract cancer, colorectal cancer, cervical cancer, thyroid cancer, salivary gland cancer, prostate cancer (e.g., hormone refractory prostate adenocarcinoma), pancreatic cancer, colon cancer, esophageal cancer, liver cancer, thyroid cancer, glioblastoma, glioma, and other neoplastic malignancies.

[0278] In some embodiments, the lung cancer is non-small cell lung cancer.

[0279] In an alternative embodiment, the lung cancer is small cell lung cancer.

[0280] In some embodiments, the lymphoma is Hodgkin's lymphoma.

[0281] In another embodiment, the lymphoma is non-Hodgkin's lymphoma. In a particular embodiment, the lymphoma is mediastinal large B-cell lymphoma.

[0282] In some embodiments, the breast cancer is triple-negative breast cancer.

[0283] In a further embodiment, the breast cancer is ER+ / HER2- breast cancer.

[0284] In some embodiments, the bladder cancer is urothelial carcinoma.

[0285] In some embodiments, the head and neck cancer is nasopharyngeal carcinoma. In some embodiments, the cancer is thyroid cancer. In other embodiments, the cancer is salivary gland cancer. In other embodiments, the cancer is squamous cell carcinoma of the head and neck.

[0286] In some embodiments, the cancer is a solid tumor with high levels of microsatellite instability (MSI-H).

[0287] In some embodiments, the cancer is a solid tumor with a high mutational burden.

[0288] In some embodiments, the cancer is metastatic colorectal cancer with high levels of microsatellite instability (MSI-H).

[0289] In some embodiments, the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma, and small cell lung cancer.

[0290] In another embodiment of the above method of treatment, the cancer is a hematological malignancy. In a particular embodiment, the hematological malignancy is acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), diffuse large B-cell lymphoma (DLBCL), EBV-positive DLBCL, primary mediastinal large B-cell lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, follicular lymphoma, Hodgkin's lymphoma (HL), mantle cell lymphoma (MCL), multiple myeloma (MM), myeloid cell leukemia 1 protein (Mcl-1), myelodysplastic syndrome (MDS), non-Hodgkin's lymphoma (NHL) or small lymphocytic lymphoma (SLL).

[0291] Malignant tumors that show improved disease-free and overall survival rates associated with the presence of tumor-infiltrating lymphocytes in biopsy or surgical specimens, such as melanoma, colorectal cancer, liver cancer, renal cancer, gastric / esophageal cancer, breast cancer, pancreatic cancer and ovarian cancer, are included in the methods and treatments described herein. Subtypes of such cancers are known to be susceptible to immune control by T lymphocytes. In addition, refractory or recurrent malignant tumors whose growth can be inhibited using the antibodies described herein are also included.

[0292] Additional cancers that may benefit from treatment with the formulations described herein include cancers associated with persistent infection with viruses such as human immunodeficiency virus, hepatitis A, B and C viruses, Epstein-Barr virus, human papilloma virus, etc., known to be causally associated with Kaposi's sarcoma, liver cancer, nasopharyngeal carcinoma, lymphoma, cervical cancer, vulvar cancer, anal cancer, penile cancer and oral cancer.

[0293] The formulations can also be used to prevent or treat infectious diseases and infectious disorders. Thus, the present invention provides a method of treating a chronic infectious disease in a mammalian subject, comprising administering to the subject an effective amount of a formulation of the present invention. In certain embodiments of the method, the formulation is administered to the subject via intravenous administration. In another embodiment, the formulation is administered to the subject by subcutaneous administration.

[0294] These agents can be used alone or in combination with vaccines to stimulate immune responses against pathogens, toxins and self-antigens. The antibodies or antigen-binding fragments thereof can be used to stimulate immune responses against viruses that infect humans, including but not limited to human immunodeficiency virus, hepatitis A, B and C viruses, Epstein-Barr virus, human cytomegalovirus, human papilloma virus and herpes viruses. Antagonist anti-PD-1 antibodies or antibody fragments can be used to stimulate immune responses against infections by bacterial or fungal parasites and other pathogens. Viral infections by hepatitis B and C viruses and HIV are considered chronic viral infections.

[0295] The formulations of the invention can be administered to a patient in combination with one or more "additional therapeutic agents." The additional therapeutic agents can be biologic therapeutic agents (including, but not limited to, antibodies against VEGF, EGFR, Her2 / neu, VEGF receptors, other growth factor receptors, CD20, CD40, CD-40L, OX-40, 4-1BB, and ICOS), immunogenic agents (e.g., attenuated cancer cells, tumor antigens, antigen-presenting cells, e.g., dendritic cells pulsed with tumor-derived antigens or nucleic acids, immunostimulatory cytokines (e.g., IL-2, IFNα2, GM-CSF), and cells transfected with genes encoding immunostimulatory cytokines (e.g., but not limited to, GM-CSF).

[0296] As mentioned above, in some embodiments of the method of the present invention, the method further comprises administering an additional therapeutic agent. In certain embodiments, the additional therapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment thereof, an anti-PD-L1 antibody or an antigen-binding fragment thereof, an anti-LAG3 antibody or an antigen-binding fragment thereof, an anti-TIGIT antibody or an antigen-binding fragment thereof, an anti-GITR antibody or an antigen-binding fragment thereof, an anti-CTL4 antibody or an antigen-binding fragment thereof, or an anti-CD27 antibody or an antigen-binding fragment thereof. In one embodiment, the additional therapeutic agent is a Newcastle Disease Virus vector expressing IL-12. In a further embodiment, the additional therapeutic agent is dinaciclib. In a further embodiment, the additional therapeutic agent is a STING agonist.

[0297] Suitable routes of administration can include, for example, parenteral delivery, e.g., intramuscular, subcutaneous and intrathecal, direct intracerebroventricular, intravenous, intraperitoneal, etc. Drugs can be administered in a variety of conventional ways, such as intraperitoneal, parenteral, intraarterial or intravenous injection. Administration methods that require limited solution volume (e.g., subcutaneous administration) require lyophilized formulations that allow reconstitution at high concentrations.

[0298] The choice of dosage of the additional therapeutic agent depends on several factors, including the serum or tissue turnover rate of the entity, the level of symptoms, the immunogenicity of the entity, and the accessibility of the target cells, tissues, or organs in the individual being treated. The dosage of the additional therapeutic agent should be an amount that produces an acceptable level of side effects. Thus, the dosage and frequency of administration of each additional therapeutic agent (e.g., biological or chemotherapeutic agent) will depend in part on the particular therapeutic agent, the severity of the cancer being treated, and the characteristics of the patient. Guidance is available for choosing appropriate dosages of antibodies, cytokines, and small molecules.See, for example: Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602; Physicians' Desk Reference 2003 (Physicians' Desk Reference, 57th Ed); Medical Economics Company; ISBN: 1563634457; 57th edition (November 2002). The determination of an appropriate dosing regimen can be made by the clinician using, for example, parameters or factors known or suspected in the art to affect or be expected to affect treatment, and depends, for example, on the patient's clinical history (e.g., previous treatments), the type and stage of the cancer being treated, and biomarkers of response to one or more therapeutic agents in the combination therapy.

[0299] To facilitate the selection of pharma- ceutically acceptable carriers or excipients for additional therapeutic agents, various references are available. See, for example: Remington's Pharmaceutical Sciences and US Pharmacopeia: National Formulary, Mack Publishing Company, Easton, PA (1984); Hardman et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY.

[0300] The pharmaceutical antibody formulations can be administered by continuous infusion or at intervals such as daily, 1-7 times per week, weekly, biweekly, 3 weeks, monthly, bimonthly, etc. Preferred dosing protocols include the maximum dose or dosing frequency that avoids significant undesirable side effects. Total weekly doses are generally at least 0.05 μg / kg, 0.2 μg / kg, 0.5 μg / kg, 1 μg / kg, 10 μg / kg, 100 μg / kg, 0.2 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 10 mg / kg, 25 mg / kg, 50 mg / kg body weight or more. See, e.g., Yang et al. (2003) New Engl. J. Med. 349:427-434; Herold et al. (2002) New Engl. J. Med. 346:1692-1698; Liu et al. (1999) J. Neurol. Neurosurg. Psych. 67:451-456; Portielji et al. (20003) Cancer Immunol. Immunother. 52:133-144. Desirable dosages of small molecule therapeutics (e.g., peptidomimetics, natural products, or organic chemicals) are approximately the same as for antibodies or polypeptides, on a "moles / kg" basis.

[0301] Embodiments of the present invention further include the use of a therapeutic agent, such as a therapeutic agent for the treatment of a human body; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; a therapeutic agent for the treatment of a human cancer; (k) reducing the severity or duration of clinical symptoms of a PD-1 associated disease or an ILT4 associated disease; (l) prolonging patient survival compared to expected survival in a similar untreated patient; (m) inducing complete or partial remission of a cancerous condition or other PD-1 associated disease or an ILT4 associated disease; (n) treating cancer; or (o) treating a chronic infection; also encompasses one or more of the biologics described herein for use in (i), for use as a medicament or composition for (ii), or for use in the preparation of a medicament for (iii).

[0302] Common methods Standard methods in molecular biology are described in: Sambrook, Fritsch and Maniatis (1982 & 1989 2 nd Edition, 2001 3 rd Edition) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, 3 rded., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard methods are also described in Ausbel, et al. (2001) Current Protocols in Molecular Biology, Vols.1-4, John Wiley and Sons, Inc. New York, NY, which describes cloning and DNA mutagenesis in bacterial cells (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugate and protein expression (Vol. 3), and bioinformatics (Vol. 4).

[0303] Protein purification methods, including immunoprecipitation, chromatography, electrophoresis, centrifugation and crystallization, have been described (Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, and glycosylation of proteins are described (see, e.g., Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York; Ausubel, et al. (2001) Current Protocols in Molecular Biology, Vol. 3, John Wiley and Sons, Inc., NY, NY, pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, MO; pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, NJ, pp. 384-391). The generation, purification and fragmentation of polyclonal and monoclonal antibodies have been described (Coligan, et al. (2001) Current Protocols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra).Standard techniques are available for characterizing ligand / receptor interactions (see, e.g., Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

[0304] Monoclonal, polyclonal and humanized antibodies can be prepared (see, e.g., Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ. Press, New York, NY; Kontermann and Dubel (eds.) (2001) Antibody Engineering, Springer-Verlag, New York; Harlow and Lane (1988) Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 139-243; Carpenter, et al. (2000) J. Immunol. 165:6205; He, et al. (1998) J. Immunol. 160:1029; Tang et al. (1999) J. Biol. Chem. 274:27371-27378; Baca et al. (1997) J. Biol. Chem. 272:10678-10684; Chothia et al. (1989) Nature 342:877-883; Foote and Winter (1992) J. Mol. Biol. 224:487-499; U.S. Patent No. 6,329,511).

[0305] An alternative to humanization is to use human antibody libraries displayed on phage or in transgenic mice (Vaughan et al. (1996) Nature Biotechnol. 14:309-314; Barbas (1995) Nature Medicine 1:837-839; Mendez et al. (1997) Nature Genetics 15:146-156; Hoogenboom and Chames (2000) Immunol. Today 21:371-377; Barbas et al. (2001) Phage Display: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Kay et al. (1996) Phage Display of Peptides and Proteins: A Laboratory Manual, Academic Press, San Diego, CA; de Bruin et al. (1999) Nature Biotechnol. 17:397-399).

[0306] It is not necessary to purify an antigen to generate antibodies. Animals can be immunized with cells bearing the antigen of interest. Splenocytes can then be isolated from the immunized animal and fused with a myeloma cell line to produce hybridomas (see, e.g., Meyaard et al. (1997) Immunity 7:283-290; Wright et al. (2000) Immunity 13:233-242; Preston et al., supra; Kaithamana et al. (1999) J. Immunol. 163:5157-5164).

[0307] Antibodies can be conjugated, for example, to small drug molecules, enzymes, liposomes, polyethylene glycol (PEG). Antibodies are useful for therapeutic, diagnostic, kits or other purposes and include, for example, antibodies conjugated with dyes, radioisotopes, enzymes or metals, such as colloidal gold (see, for example, Le Doussal et al. (1991) J. Immunol. 146:169-175; Gibellini et al. (1998) J. Immunol. 160:3891-3898; Hsing and Bishop (1999) J. Immunol. 162:2804-2811; Everts et al. (2002) J. Immunol. 168:883-889).

[0308] Methods for flow cytometry, including fluorescence-activated cell sorting (FACS), are available (see, e.g., Owens, et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, 2 nd ed.; Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies, for use, e.g., as diagnostic reagents, are available (Molecular Probesy (2003) Catalogue, Molecular Probes, Inc., Eugene, OR; Sigma-Aldrich (2003) Catalogue, St. Louis, MO).

[0309] Standard methods for immune system histology have been described (see, e.g., Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott, Williams, and Wilkins, Phila, PA; Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

[0310] Software packages and databases are available for determining, for example, antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments (see, e.g., GenBank, Vector NTI® Suite (Informax, Inc., Bethesda, MD); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DeCypher® (TimeLogic Corp., Crystal Bay, Nevada); Menne, et al. (2000) Bioinformatics 16: 741-742; Menne, et al. (2000) Bioinformatics Applications Note 16:741-742; Wren, et al. (2002) Comput. Methods Programs Biomed. 68:177-181; von Heijne (1983) Eur. J. Biochem. 133:17-21; von Heijne (1986) Nucleic Acids Res. 14:4683-4690).

[0311] Analysis method Suitable analytical methods for evaluating the stability of a product include size exclusion chromatography (SEC), dynamic light scattering (DLS), differential scanning calorimetry (DSC), iso-asp quantification, titer, UV at 340 nm, UV spectroscopy and FTIR. SEC (J. Pharm. Scien., 83:1645-1650, (1994); Pharm. Res., 11:485 (1994); J. Pharm. Bio. Anal., 15:1928 (1997); J. Pharm. Bio. Anal., 14:1133-1140 (1986)) measures the percentage of monomer in the product and provides information on the amount of soluble aggregates. DSC (Pharm. Res., 15:200 (1998); Pharm. Res., 9:109 (1982)) provides information on the denaturation temperature and glass transition temperature of the protein. DLS (American Lab., November (1991)) measures the average diffusion coefficient and gives information on the amount of soluble and insoluble aggregates. UV at 340 nm measures the scattered light intensity at 340 nm and gives information on the amount of soluble and insoluble aggregates. UV spectroscopy measures the absorbance at 278 nm and gives information on the protein concentration. FTIR (Eur. J. Pharm. Biopharm., 45:231 (1998); Pharm. Res., 12:1250 (1995); J. Pharm. Scien., 85:1290 (1996); J. Pharm. Scien., 87:1069 (1998)) measures the IR spectrum in the amide 1 region and gives information on the secondary structure of the protein.

[0312] The iso-asp content in the samples is measured using the Isoquant Isoaspartate Detection System (Promega). This kit uses the enzyme protein isoaspartyl methyltransferase (PIMT) to specifically detect the presence of isoaspartic acid residues in target proteins. PIMT catalyzes the transfer of a methyl group at the α-carboxyl position from S-adenosyl-L-methionine to isoaspartic acid, generating S-adenosyl-L-homocysteine ​​(SAH) in the process. This is a relatively small molecule that can usually be isolated and quantified by reversed-phase HPLC using the SAH HPLC standard that is included in the kit.

[0313] The potency or bioidentity of an antibody can be measured by its ability to bind to its antigen. Specific binding of an antibody to its antigen can be quantified by any method known to those skilled in the art, for example by immunoassays such as ELISA (enzyme-linked immunosorbent assay).

[0314] All publications mentioned herein are incorporated by reference for the purpose of describing and disclosing methodologies and materials that might be used in connection with the present invention.

[0315] Although various embodiments of the present invention have been described herein with reference to the accompanying drawings, it is to be understood that the invention is not limited to these precise embodiments, and various changes and modifications may be made by those skilled in the art without departing from the scope or spirit of the present invention as defined in the appended claims.

[0316] Embodiment 1 provides a formulation of an anti-human immunoglobulin-like transcript 4 ("anti-ILT4") antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM of a buffer; (iii) about 6% to about 8% weight / volume (w / v) of a non-reducing sugar; (iv) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (v) about 1 mM to about 20 mM of an antioxidant; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has the following structure: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17). and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); wherein the buffer is an L-histidine buffer, an acetate buffer or a citrate buffer, wherein the non-reducing sugar is a disaccharide, wherein the non-ionic surfactant is polysorbate 20 or polysorbate 80, and wherein the antioxidant is methionine.

[0317] Embodiment 2 provides the formulation of embodiment 1, wherein the buffer is an L-histidine buffer.

[0318] Embodiment 3 provides the formulation of embodiment 1 or 2, wherein the non-reducing sugar is sucrose.

[0319] Embodiment 4 provides a formulation of any one of embodiments 1-3, wherein the non-ionic surfactant is polysorbate 80.

[0320] Embodiment 5 provides the formulation of any one of embodiments 1-4, wherein the antioxidant is L-methionine.

[0321] Embodiment 6 provides a formulation of an anti-human immunoglobulin-like transcript 4 ("anti-ILT4") antibody or antigen-binding fragment thereof comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM L-histidine buffer; (iii) about 6% to about 8% weight / volume (w / v) sucrose; (iv) about 0.01% to about 0.10% (w / v) polysorbate 80; and (v) about 1 mM to about 20 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a CDR- a heavy chain variable domain comprising H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0322] Embodiment 7 provides the formulation of embodiment 6 comprising about 8 mM to about 12 mM L-histidine buffer.

[0323] Embodiment 8 provides a formulation of embodiment 6 or 7 comprising about 5 mM to about 10 mM L-methionine.

[0324] Embodiment 9 provides a formulation according to any of embodiments 6 to 8, comprising about 0.01% to about 0.05% (w / v) polysorbate 80.

[0325] Embodiment 10 provides the formulation of any of Embodiments 1-9 comprising about 10 mg / mL to about 150 mg / mL of said anti-ILT4 antibody or antigen-binding fragment thereof.

[0326] Embodiment 11 provides the formulation of embodiment 10, wherein the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL or about 150 mg / mL.

[0327] Embodiment 12 provides a formulation of any of embodiments 1-11 comprising about 25 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0328] Embodiment 13 provides a formulation of any of embodiments 1-11 comprising about 50 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0329] Embodiment 14 provides a formulation of any of embodiments 1-11 comprising about 75 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0330] Embodiment 15 provides a formulation of any of embodiments 1-11 comprising about 100 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0331] Embodiment 16 provides a formulation of any of embodiments 1-11, comprising about 125 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

[0332] Embodiment 17 provides a formulation according to any one of embodiments 1 to 16, wherein the formulation has a pH of about 5.0 to about 6.8.

[0333] Embodiment 18 provides a formulation of any one of embodiments 1 to 16, wherein the formulation has a pH of about 5.5 to about 6.0.

[0334] Embodiment 19 provides a formulation of any one of embodiments 1-16, wherein the formulation has a pH of about 5.5.

[0335]

[0033] Embodiment 20 provides a formulation of an anti-human immunoglobulin-like transcript 4 ("anti-ILT4") antibody or antigen-binding fragment thereof comprising: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer, pH about 5.5; (iii) about 7% weight / volume (w / v) sucrose; (iv) about 0.025% (w / v) polysorbate 80; and (v) about 10 mM L-methionine; wherein the anti-ILT4 antibody or antigen-binding fragment thereof has CDR-H1:GYY. WS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17) and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20) and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0336] Embodiment 21 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0337] Embodiment 22 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

[0338] Embodiment 23 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:58.

[0339] Embodiment 24 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:7.

[0340] Embodiment 25 provides the formulation of any of the embodiments of claims 1 to 20, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 80; and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 7.

[0341] Embodiment 26 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO: 58.

[0342] Embodiment 27 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:7.

[0343] Embodiment 28 provides the formulation of any one of Embodiments 1 to 20, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 80; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 7.

[0344] Embodiment 29 provides the formulation of any one of embodiments 1 to 28, wherein the formulation is a liquid formulation stored at about 3° C. to about 5° C., frozen at or below −20° C., frozen at or below −70° C., or a reconstituted solution from a lyophilized formulation.

[0345] Embodiment 30 is a method for producing a composition comprising, after storage for 6 months at about 3° C. to about 5° C.: (i) the % monomer of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) the OD 350-500(iii) the turbidity of the formulation is at most about 0.135 when measured by high performance ion exchange chromatography; (iv) the number of subvisible particles of particles at least 2 μm in size is at most about 3500 when measured by microflow imaging; and / or (v) the % oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359 and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% when measured by reduced peptide mapping;

[0346] Embodiment 31 provides the formulation of any of embodiments 1-30, wherein the anti-ILT4 antibody, or antigen-binding fragment thereof, is a monoclonal antibody.

[0347] Embodiment 32 provides a method of treating cancer in a human patient in need thereof, comprising administering an effective amount of the formulation of any one of embodiments 1 to 31.

[0348] Embodiment 33 provides the method of embodiment 32, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma, and small cell lung cancer.

[0349] Embodiment 34 provides the use of a formulation according to any one of embodiments 1 to 31 for the preparation of a medicament for treating cancer.

[0350] Embodiment 35 provides the use of a formulation according to any one of embodiments 1 to 31 for treating cancer in a human patient.

[0351] Embodiment 36 provides the use according to embodiment 34, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma and small cell lung cancer. EXAMPLES

[0352] Example 1 Anti-ILT4 preformulation analysis The pre-formulation development feasibility plan is summarized in Table 3. Fully human anti-ILT4 monoclonal antibody 1E1 ("Antibody 1") with a human lambda light chain constant domain and a human IgG4-S228P heavy chain constant domain was tested. All formulations were prepared in 96-well Axygen polypropylene deep well plates (product no. P-DW-11-C) using a JANUS automated liquid handling system equipped with a Parisian pipetting arm. In each well was a 500 μL aliquot with Antibody 1 at a final concentration of 1 mg / mL. [Table 4]

[0353] 200 μL aliquots were removed from the plate for initial sample analysis (20 μL for differential scanning fluorimetry (DSF) thermal transition analysis and 180 μL for subvisible particle analysis using a Guava® easyCyte flow cytometer). The remaining 300 μL of sample in the 96-well Axygen plate was analyzed using a Nunc TMThe plates were covered with a Microplate Sealing Cap Mat (product no. 276000) and tightly sealed. The plates were double-wrapped in SCS Dri-Shield® 2000 Series Moisture Barrier Bags (product no. 700810) to prevent evaporation. The sealed plates were stressed in a thermal chamber at 50° C. for 10 days. Protein concentration was monitored before and after heat stress to account for any increase in protein concentration due to evaporation. The increase in protein concentration across the plate was less than 5%. All standard precautions were observed when handling protein solutions (e.g., equilibrating the solution to room temperature, minimal shaking, and exposure to room light).

[0354] After heat stress, the plate was equilibrated at room temperature for 30 minutes. The plate was then carefully opened in a laminar flow hood. The stressed samples were mixed up to 5 times using a 200 μL multichannel pipette. 200 μL of the sample was then removed and dispensed into a 96-well Costar Clear Plate (product no. 3635) and absorbance was measured at wavelengths of 280, 320, 350 and 500 nm. After UV analysis, 180 μL of the 200 μL aliquot was carefully transferred to a Corning® 96-well Clear V-bottom Microplate (product no. 3894) for analysis of particles subvisible to the naked eye. The remaining 20 μL was used for quantitative protein purity analysis by reduced high-throughput CE-SDS using Caliper's LabChip GXII system. On the same day, 100 μL of the stressed samples remaining in the Axygen deep well plate were transferred to an ABgene 96-well PCR plate (product no. AB-2800) along with the initial (unstressed) samples for UP-SEC and cIEF analysis.

[0355] Thermal stability with DSF The thermal stability of Antibody 1 formulations was evaluated by DSF under various buffer, pH and ionic strength conditions. The Tm1 (first unfolding midpoint) in most tested conditions was in the range of 60-66°C (data not shown). Overall, the results of this assay showed no adverse effects in terms of thermal stability.

[0356] Soluble aggregates by UP-SEC UP-SEC analysis of stressed samples of Antibody 1 in various formulation conditions showed similar colloidal stability profiles (data not shown). Antibody 1 formulations showed resistance to fragmentation and showed greater than 3% soluble aggregation in formulations with a pH below 5.5 (data not shown).

[0357] Subvisible Particles by Guava® easyCyte Flow Cytometer Sub-visible particle (0.22 microns to 10 microns) testing was performed on a Guava® easyCyte flow cytometer for Antibody 1. Antibody 1 did not show significant formation of sub-visible aggregates after stress in most formulation compositions (data not shown).

[0358] Fragmentation by CE-SDS To comprehensively understand the proteolytic cleavage of the heavy or light chains of the anti-ILT4 monoclonal antibody, the stressed samples were also analyzed by reducing sodium dodecyl sulfate capillary electrophoresis (R-CE-SDS). Fragmentation of Antibody 1 was minimal in 10 mM histidine at pH range 5.6-6.8 or in 10 mM acetate at pH range 5.0-5.6.

[0359] Chemical stability by cIEF The chemical stability of Antibody 1 formulations was assessed by cIEF under various buffer, pH and ionic strength conditions. cIEF profiles showed a decrease in the relative main peak area and a significant increase in the relative peak area of ​​the acidic variant after 10 days at 50°C. The addition of salt did not result in significant improvement in protein stability across the compositions examined. The formulations giving the greatest stability to Antibody 1 were identified as 10 mM histidine in the pH range of 5.5-6.0 or 10 mM acetate at pH 5.6.

[0360] The stressed samples were also subjected to turbidity analysis by UV / Vis spectrophotometer to evaluate colloidal instability (A 350-500 ) The results showed no trends as a function of any of the parameters evaluated.

[0361] The results above demonstrated that formulations of Antibody 1 had good colloidal stability profiles (measured by turbidity and subvisible particle analysis) and thermal stability profiles, and that the unformulated antibody exhibited chemical instability and a tendency to aggregate. As a result, buffers of 10 mM histidine or 10 mM acetate in the pH range of about 5.5-6.0 were selected for further formulation development.

[0362] Example 2 Early feasibility assessment of anti-ILT4 agents Five anti-ILT4 monoclonal antibody formulations were prepared at a concentration of 50 mg / mL of Antibody 1 in the following matrices: 10 mM acetate, pH 5.4; 10 mM acetate, pH 5.8; 10 mM L-histidine, pH 5.4; 10 mM L-histidine, pH 5.8; and 10 mM L-histidine, pH 5.8, with 9% (w / v) sucrose. The concentrateability of the Antibody 1 formulations was further evaluated by concentrating Antibody 1 to 100 mg / mL in each of the five formulation matrices. The formulations were analyzed for appearance, turbidity (OD 350-500 ), viscosity, concentration and pH. The diffusion interaction parameter (k D) and relative solubility values ​​to determine the self-association of Antibody 1 in each of the five formulations.

[0363] The data are summarized in Table 4. From this data, the colloidal stability (OD 350-500 ) was superior at low pH (5.4) than at high pH (5.8) in both 10 mM acetate and 10 mM histidine buffers, suggesting improved stability in the presence of 9% sucrose as a stabilizer (10 mM histidine buffer, pH 5.8). Overall, k D The value of k was negative, suggesting an inherent tendency of antibodies to self-associate. D The values ​​of σ were less negative at low pH (5.4) in both buffers, suggesting less self-association at this pH. The presence of the stabilizer sucrose was found to reduce the crowding effect in 10 mM histidine buffer at pH 5.8. Overall, the relative solubility of Antibody 1 in 10 mM acetate buffers (pH 5.4 and 5.8) and 10 mM histidine buffer (pH 5.4) was comparable. Within the 10 mM histidine buffer, the solubility was relatively lower at the higher pH (5.8). The viscosity of the five formulations at a concentration of 100 mg / mL (similar values) was not affected by pH or formulation buffer, indicating the ability of the molecule to concentrate beyond 50 mg / mL.

[0364] [Table 5]

[0365] Example 3 Forced degradation test of anti-ILT4 formulation Forced degradation studies were performed on formulations containing Antibody 1. The Antibody 1 formulations tested contained 50 mg / mL antibody, 10 mM L-histidine buffer (pH 6.0), 7% (w / v) sucrose and 0.02% (w / v) PS-80. Antibody 1 formulations were exposed to a variety of stress conditions including heat (50°C, up to 7 days), high pH (pH 10, 25°C, up to 7 days), low pH (pH 3.5, 25°C, up to 7 days), light (25°C, up to 2x ICH scale) or peroxide (0.1% tBHP, 25°C, up to 24 hours).

[0366] Stressed samples were visually inspected for color and particulates and examined by UV, UP-SEC, NR-CE-SDS, R-CE-SDS, HP-IEX, DLS and LC-MS (intact and reduced mass and reduced peptide mapping).

[0367] Major degradation pathways (>10% change at end of stress compared to initial point) included: increase in acidic variants due to heat (50°C), high pH (pH 10.0) and light exposure stresses; increase in deamidation and isomerization due to high pH stress; increase in HMW species, basic variants and Trp oxidation due to light exposure (2-fold) stress; and increase in Met oxidation due to light exposure and peroxide (0.1% tBHP) stresses. No major degradation pathways were observed with low pH (pH 3.5) stress.

[0368] Minor degradation (less than 10% change at the end of stress compared to the initial point) included: increase in HMW species due to heat, high pH and low pH stress; increase in LMW species due to heat, high pH, ​​low pH and light exposure stress; increase in turbidity due to light exposure stress; and increase in basic mutants due to peroxide stress.

[0369] The results of the light stress studies are summarized below in Table 5. Detailed results of the other stress studies are not shown. [Table 6]

[0370] The effect of light exposure on potency at 2x ICH scale was detected by ELISA. GeoMean values ​​were 106 and 28 for dark control and 2x exposure stress samples, respectively, suggesting that light exposure reduces the potency of Antibody 1.

[0371] Example 4 Early stability studies of anti-ILT4 formulations Early feasibility data showed that a lower pH (5.4±0.2) was preferred based on good colloidal, biophysical and chemical stability (Example 2). Based on forced degradation studies (Example 3), photo-oxidation was known to be one of the major hurdles for anti-ILT4 monoclonal antibodies. Four different formulations of antibody 1 were staged for early thermal stability at a target pH of 5.5 and an antibody concentration of 50 mg / mL. Two formulation matrices consisted of 10 mM histidine (with or without 5 mM L-methionine as an antioxidant) and the remaining two consisted of 10 mM acetate (with or without 5 mM L-methionine as an antioxidant); each of the four formulations was prepared at pH 5.5 with 7% (w / v) sucrose as a stabilizer and 0.025% (w / v) polysorbate 80 (PS-80) as a surfactant. The 10 mM L-histidine buffer consisted of 0.29 mg / mL L-histidine and 1.71 mg / mL L-histidine monohydrochloride monohydrate with a target formulation pH of 5.5. 10 mM L-methionine in the formulation corresponds to 1.49 mg / mL L-methionine, 7% (w / v) sucrose and 0.025% (w / v) polysorbate 80 correspond to 70 mg / mL sucrose and 0.25 mg / mL PS-80 in the formulation, respectively. Formulations were staged for stability up to 6 months under ICH conditions: 5° C. (5° C.±3° C.), 25° C. (25° C., 60% RH), and 40° C. (40° C., 75% RH). The stability plan is shown in Table 6 below. Four pairs of formulations were analyzed for appearance, pH, protein concentration (UV 280 ), turbidity (OD 350-500The samples were tested for colloidal stability by HPLC (HPLC-HPLC), purity by ultra-performance size-exclusion chromatography (UP-SEC), charge variants by high performance ion-exchange chromatography (HP-IEX), number of subvisible particles by microflow imaging (MFI), and % change in oxidation.

[0372] [Table 7]

[0373] Early stability data for the four anti-ILT4 monoclonal antibody formulations are summarized in Tables 7-10. Stability trends for the four formulations are shown in Figures 1A-1I and 2A-2I. As seen in these figures, Antibody 1 showed similar stability profiles at all three stability conditions (5°C, 25°C, 40°C) in the presence of 7% (w / v) sucrose as a stabilizer and 0.025% (w / v) polysorbate 80 as a surfactant in both acetate and L-histidine buffers at pH 5.5. In all assays, the degradation rates were higher under accelerated conditions at 25°C and even higher under stressed conditions at 40°C. Antibody 1 formulations were stable up to 6 months at 5°C storage according to the stability plan. The presence of 5 mM L-methionine as an antioxidant in the formulation improved colloidal stability (OD) to some extent for anti-ILT4 monoclonal antibody formulations in two buffer matrices under accelerated conditions at 25°C and stressed conditions at 40°C. 350-500) and reduced soluble aggregates (% HMW) (Figure 1A-Figure 1I). 5 mM L-methionine did not affect the charge profile by HP-IEX (Figure 2A-Figure 2I). L-histidine was selected as the lead buffer and acetate was selected as the backup buffer. As seen in the reduced peptide mapping results in Tables 8-11, the presence of 5 mM L-methionine in either acetate or L-histidine formulations was found to reduce the oxidation levels of methionine residues (M359, M253, and M429) and, to some extent, tryptophan residues (W102 and W7) due to heat stress, especially in the case of the L-histidine formulation.

[0374] [Table 8]

[0375] [Table 9]

[0376] [Table 10]

[0377] [Table 11]

[0378] Example 5 pH range stability study of anti-ILT4 formulations The pH ranging study was conducted to measure the drug product stability of Antibody 1 at ±0.5 pH units from the target formulation pH of 5.5, i.e., at pH 5.0 or pH 6.0, and to help define the pH specification for formal stability. Four pH ranging formulations of Antibody 1 were staged for stability under ICH conditions (5°C (5°C ±3°C), 25°C (25°C, 60% RH), and 40°C (40°C, 75% RH)) for 6 months. Two formulations were prepared in 10 mM histidine buffer with 7% (w / v) sucrose as a stabilizer and 0.025% (w / v) polysorbate 80 as a surfactant; one formulation at pH 5.0 and the other at pH 6.0. Two formulations were prepared in 10 mM acetate buffer with 7% (w / v) sucrose as a stabilizer and 0.025% (w / v) polysorbate 80 as a surfactant; one formulation at pH 5.0 and the other at pH 6.0. Purified antibody 1 was dialyzed against each buffer and the pH of the final formulation was measured after dialysis, no pH adjustment was performed after this step. The final formulated drug substance was filtered to obtain the bulk of the drug product, which was then filled into 2 mL glass vials and sealed with rubber stoppers. The stability plan was the same as shown in Table 6. The four formulations were analyzed for appearance, pH, protein concentration (UV 280 ), turbidity (OD 350-500 The samples were tested for colloidal stability by HPLC-MS / MSI measurement, purity by UP-SEC, charge variants by HP-IEX and number of subvisible particles by MFI.

[0379] The data are summarized in Tables 11-14. The stability trends for the four antibody 1 pH ranging formulations are shown in Figures 3A-3I and 4A-4I, with data from an early stability study at pH 5.5 included for comparison. As can be seen in these figures, there was no significant change in the anti-ILT4 monoclonal antibody formulations in acetate or histidine buffer matrices at 5°C storage at either extreme pH condition (pH 5.0 and pH 6.0) compared to the target pH of 5.5. The anti-ILT4 monoclonal antibody was found to be stable within ±0.5 units of the target pH of 5.5. Under accelerated conditions at 25°C and stress conditions at 40°C, the anti-ILT4 monoclonal antibody showed better colloidal stability (OD 2.0) at low pH (5.0), then at the target pH (5.5) and at high pH (6.0). 350-500 ), physical stability (% HMW and % monomer) and chemical stability. Based on these data sets, pH 5.5 was selected as the pH for the anti-ILT4 monoclonal antibody formulation.

[0380] [Table 12]

[0381] [Table 13]

[0382] [Table 14]

[0383] [Table 15]

[0384] Example 6 Anti-ILT4 formulation surfactant ranging study A detergent ranging study of anti-ILT4 monoclonal antibody formulations containing Antibody 1 was performed with polysorbate 80 (PS-80) to analyze the sensitivity of the antibody to agitation stress in the presence or absence of detergent. The anti-ILT4 monoclonal antibody formulation consisted of 50 mg / mL of Antibody 1 in 10 mM histidine buffer (pH 5.5) and 7% (w / v) sucrose. Five different formulations were prepared, each containing a range of polysorbate 80 (i.e., 0% (w / v) or 0 mg / mL PS-80, 0.01% (w / v) or 0.1 mg / mL PS-80, 0.025% (w / v) or 0.25 mg / mL PS-80, 0.05% (w / v) or 0.5 mg / mL PS-80, and 0.1% (w / v) or 1 mg / mL PS-80). 2 mL glass vials were filled with 2.2 mL of filtered anti-ILT4 bulk drug product and sealed with rubber stoppers. Anti-ILT4 drug product vials were agitated horizontally (ambient conditions) at 300 rpm on a Thermo Scientific Shaker for up to 3 days or up to 7 days. To eliminate temperature effects, the same five anti-ILT4 formulations (PS-80 ranging) were left at room temperature without shaking for 7 days and used as controls for each study. All vials (stress and ambient controls) were covered with aluminum foil to eliminate light effects. Samples were analyzed for appearance, protein concentration (UV 280 ), turbidity (OD 350-500 ), sub-visible particulates by MFI, purity by UP-SEC, and charge variants by HP-IEX.

[0385] The data are summarized in Figures 7 and 8. The stability trends of the five anti-ILT4 monoclonal antibody surfactant blend formulations are shown in Figures 5A-5G. As can be seen in these figures, in the absence of surfactant, visible particles were generated with increasing amounts of stirring from 3 to 7 days. The ambient control samples at 7 days also showed very little visible particles. Turbidity (OD 350-500), an increase in the number of subvisible particles (≥ 2 μm, ≥ 5 μm, ≥ 10 μm, and ≥ 25 μm), and an increase in the soluble aggregate level (% HMW) were observed. In the presence of 0.1 mg / mL [0.01% (w / v)], 0.25 mg / mL [0.025% (w / v)], 0.5 mg / mL [0.5% (w / v)], or 1.0 mg / mL [0.1% (w / v)] of surfactant, the appearance, turbidity (OD 350-500 No significant differences were observed in the solubility (%) or soluble aggregate levels (% HMW). Concentrations of PS-80 of 0.5 mg / mL and above resulted in some increase in subvisible particles up to 7 days of stirring.

[0386] The data suggest that the surfactant plays an important role in preventing agitation-induced aggregation. All stability attributes were comparable after the addition of surfactant. Overall, 0.25 mg / mL PS-80 appeared to minimize particle content compared to 0.5 mg / mL or 1 mg / mL PS-80. Thus, no significant advantage was gained from adding higher levels of surfactant. As a result, a PS-80 concentration of 0.25 mg / mL was chosen for the L-histidine formulation containing 50 mg / mL anti-ILT4 monoclonal antibody.

[0387] Example 7 Light stress study of anti-ILT4 formulations with 10 mM L-methionine During forced degradation studies, the major degradation pathway of anti-ILT4 monoclonal antibody was identified as light-induced oxidation, affecting the potency of anti-ILT4 monoclonal antibody (Example 3). Early stability data (Example 4) suggested that 5 mM L-methionine was effective in reducing oxidation of L-methionine and L-tryptophan residues due to heat stress. To further explore the effect of higher L-methionine content (10 mM) in anti-ILT4 monoclonal antibody L-histidine formulations, light stress studies were performed.

[0388] Based on early stability data, 10 mM L-histidine, pH 5.5 was finalized as the buffer-pH matrix for the formulation development activities. Two L-histidine formulations were tested. One formulation (H) contained 50 mg / mL anti-ILT4 monoclonal antibody, 10 mM L-histidine, pH 5.5, 7% (w / v) sucrose, and 0.025% (w / v) polysorbate 80; the other formulation (H+M) contained 50 mg / mL anti-ILT4 monoclonal antibody, 10 mM L-histidine, pH 5.5, 10 mM L-methionine, 7% (w / v) sucrose, and 0.025% (w / v) polysorbate 80. The third formulation (HC) consisted of 50 mg / mL anti-ILT4 monoclonal antibody, 10 mM L-histidine (pH 5.5), 7% (w / v) sucrose, and 0.025% (w / v) polysorbate 80 sucrose, and was subjected to 0.2 ICH, 0.5 ICH, or 1 ICH of light stress (ultraviolet light-UV and cool white light-CWL, or a combination of visible light) in Caron Photostability Chambers at room temperature, as well as dark control (non-light exposed). To eliminate the effect of temperature, the dark control samples were covered with aluminum foil and placed in the same chamber as the light-exposed samples. Samples were exposed to a cumulative amount of light exposure and removed when the exposure limit was reached. Samples were analyzed for appearance, protein concentration (UV, CWL, or visible light), and protein concentration (P < 0.05). 280 ), turbidity (OD 350-500 ), purity by UP-SEC, charge variants by HP-IEX, oxidation level by reduced peptide mapping (anti-ILT4 monoclonal antibody oxidation hotspots M359, W7, M253, W102, M429) and potency by binding ELISA.

[0389] As seen in Figures 6A-6K and 9, 10 mM L-methionine significantly increased colloidal stability (OD 350-500It was found that the efficacy of the 10 mM L-methionine was improved against extreme light conditions (1ICH) in the presence of 10 mM L-methionine, as shown in FIG. 9. Thus, the lead formulation of anti-ILT4 monoclonal antibody was selected as follows: 50 mg / mL anti-ILT4 antibody, 0.29 mg / mL L-histidine, 1.71 mg / mL L-histidine monohydrochloride monohydrate, 1.49 mg / mL L-methionine, 70 mg / mL sucrose, and 0.25 mg / mL polysorbate 80.

[0390] Example 8 Anti-ILT4 formulation freeze-thaw study The stability of anti-ILT4 monoclonal antibody lead formulation (50 mg / mL anti-ILT4, 0.29 mg / mL L-histidine, 1.71 mg / mL L-histidine monohydrochloride monohydrate, 1.49 mg / mL L-methionine, 70 mg / mL sucrose, 0.25 mg / mL polysorbate 80, pH 5.5) was evaluated under freeze-thaw stress. Anti-ILT4 drug product was filled into 2 mL type 1 glass vials using 2.2 mL volume and sealed with rubber stoppers. The drug product vials were subjected to up to five freeze-thaw cycles (i.e., 1×F / T, 3×F / T, and 5×F / T); each cycle consisted of freezing at -80°C followed by thawing at 25°C (25°C, 60% RH chamber) with a 5°C control. Samples were analyzed for appearance, protein concentration (UV 280 ), turbidity (OD 350-500 ), purity by UP-SEC, and charge variants by HP-IEX.

[0391] As shown in Table 15, the anti-ILT4 L-histidine lead formulation showed no change up to five freeze-thaw cycles. Appearance, turbidity (OD 350-500), subvisible particle numbers and biochemical profiles (UP-SEC and HP-IEX) were comparable up to 5-fold freeze-thaw stress. Overall, freeze-thaw cycling had no measurable effect on aggregation or biochemical properties of the anti-ILT4 lead formulation.

[0392] Example 9 Characteristics of anti-ILT4 drugs The properties of the anti-ILT4 monoclonal antibody formulations, such as density, viscosity, osmolality and apparent glass transition temperature (Tg'), were measured.

[0393] Apparent glass transition temperature (T)g' Modulated differential scanning microcalorimetry (mDSC) was used to measure Tg'. The following parameters were used during the run: temperature equilibrated at -90.0°C; modulation of ±0.5°C every 60 seconds; isothermal for 5.0 minutes, ramped at 2.0°C / min to 25.0°C.

[0394] Osmolality The Advanced Instruments, Inc. osmometer was calibrated using two calibration standards (100 mOsm / kg and 500 mOsm / kg) provided by Advanced Instruments, Inc., and was measured to be 100 mOsm / kg and 500 mOsm / kg, respectively (within the acceptable range). 250 μL of sample was dispensed into the sample dispenser, and the osmolality was measured in triplicate.

[0395] density An Anton Paar Densitometer was used for density measurements. The vibrating U-tube of the density meter was washed with water, then with ethanol, and air-dried with the built-in pump. Prior to the density measurement of the samples, the air density in the vibrating U-tube was 0.00126 g / cm at 20°C. 3The density was measured to be 0.05 to 0.05%. For density measurements, 1 mL of sample was loaded into a sterile 1 mL syringe. The sample was ensured to be bubble-free and gently inserted into the vibrating U-tube through the sample port. Density measurements were performed at 5°C, 20°C, and 30°C.

[0396] viscosity A Rheosense mVROC viscometer was used for viscosity measurements. The water bath was stabilized at 20°C and a system check was performed with aquet solution. Approximately 400 μL of sample was taken into a clean 0.5 mL syringe and fixed into the system for measurement. Viscosity was measured at 20°C using a flow rate of 50 μL / min with a measurement time of 60 seconds. Viscosity values ​​with a slope-fitted Rsqrd value of at least 0.98 were used for recording.

[0397] The properties of the anti-ILT4 monoclonal antibody formulation (100 mg / vial (50 mg / mL)), such as apparent glass transition temperature (Tg'), osmolality, density, viscosity and extractable volume, are summarized in Table 16. [Table 16]

[0398] Example 10: Ion exchange (IEX) method for measuring acidic species of anti-ILT4 antibodies For the IEX method, a Waters Alliance LC system (Milford, MA, USA) was used, with a Thermo Scientific's ProPac WCX-10 (p / n:054993, particle size 10um, diameter 4mm, length 250mm) selected with a loading of 80μg sample. A mobile phase (A) 24mM MES (pH 6.1) (containing 4% acetonitrile) and a mobile phase (B) 20mM sodium phosphate, 95mM NaCl (pH 8.0) (containing 4% acetonitrile) were used as a nonlinear sigmoidal pH gradient, and the separation was monitored over 34 minutes at a flow rate of 0.5mL / min and a column temperature of 35°C. The gradient used was as follows: 0-0.6 min 22%-22% B; 0.6-15.0 min 22%-29% B; 15.0-30.0 min 29%-70% B; 30.0-30.5 min 70%-100% B; and 30.5-33.0 min 100%-100% B. The column was stripped with mobile phase (C) (10 mM CHES (pH 8.0), 40 mM Tris, 15 mM EDTA, 200 mM NaCl, and 4% acetonitrile) at 0.5 mL / min from 33.1-34.0 min, followed by re-equilibration at 1.0 mL / min with 22% B from 34.5-44.5 min. The flow rate was reduced to 0.5 mL / min from 44.5-45 min. The eluate was monitored at 280 nm for peak detection, and the assay variability was determined to be within 1%.

[0399] [Table 17]

Claims

1. 1. A formulation of an anti-human immunoglobulin-like transcript 4 (anti-ILT4) antibody or antigen-binding fragment thereof, comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM of a buffering agent; (iii) about 6% to about 8% weight / volume (w / v) non-reducing sugars; (iv) about 0.01% to about 0.10% (w / v) of a non-ionic surfactant; and (v) about 1 mM to about 20 mM of an antioxidant; Including; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); wherein the buffer is an L-histidine buffer, an acetate buffer, or a citrate buffer; wherein the non-reducing sugar is a disaccharide; wherein the non-ionic surfactant is polysorbate 20 or polysorbate 80; and wherein the antioxidant is methionine; The formulation.

2. 2. The formulation of claim 1, wherein the buffer is an L-histidine buffer.

3. 3. The formulation of claim 1 or 2, wherein the non-reducing sugar is sucrose.

4. The formulation of any one of claims 1 to 3, wherein the non-ionic surfactant is polysorbate 80.

5. The formulation according to any one of claims 1 to 4, wherein the antioxidant is L-methionine.

6. 1. A formulation of an anti-human immunoglobulin-like transcript 4 (anti-ILT4) antibody or antigen-binding fragment thereof, comprising: (i) about 10 mg / mL to about 200 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 5 mM to about 20 mM L-histidine buffer; (iii) about 6% to about 8% weight / volume (w / v) sucrose; (iv) about 0.01% to about 0.10% (w / v) polysorbate 80; and (v) about 1 mM to about 20 mM L-methionine; Including; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); The formulation.

7. 7. The formulation of claim 6, comprising about 8 mM to about 12 mM L-histidine buffer.

8. 8. The formulation of claim 6 or 7, comprising about 5 mM to about 10 mM L-methionine.

9. The formulation of any one of claims 6 to 8, comprising about 0.01% to about 0.05% (w / v) polysorbate 80.

10. The formulation of any of claims 1-9, comprising about 10 mg / mL to about 150 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof.

11. 11. The formulation of claim 10, wherein the concentration of the anti-ILT4 antibody or antigen-binding fragment thereof is about 10 mg / mL, about 12.5 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL or about 150 mg / mL.

12. 12. The formulation of any of claims 1-11, comprising about 25 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

13. 12. The formulation of any of claims 1-11, comprising about 50 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

14. 12. The formulation of any of claims 1-11, comprising about 75 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

15. 12. The formulation of any of claims 1-11, comprising about 100 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

16. 12. The formulation of any of claims 1-11, comprising about 125 mg / mL of the anti-ILT4 antibody or antigen-binding fragment thereof, about 10 mM L-histidine buffer, about 7% w / v sucrose, about 0.025% polysorbate 80, and about 10 mM L-methionine.

17. The formulation of any one of claims 1 to 16, wherein the formulation has a pH of about 5.0 to about 6.

8.

18. The formulation of any one of claims 1 to 16, wherein the formulation has a pH of about 5.5 to about 6.

0.

19. The formulation of any one of claims 1 to 16, wherein the formulation has a pH of about 5.

5.

20. 1. A formulation of an anti-human immunoglobulin-like transcript 4 (anti-ILT4) antibody or antigen-binding fragment thereof, comprising: (i) about 50 mg / mL to about 100 mg / mL of an anti-ILT4 antibody or antigen-binding fragment thereof; (ii) about 10 mM L-histidine buffer, pH about 5.5; (iii) about 7% weight / volume (w / v) sucrose; (iv) about 0.025% (w / v) polysorbate 80; and (v) about 10 mM L-methionine; Including; wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHXGSTNYNPSLKS (wherein X is S or A) (SEQ ID NO: 17), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E, or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21); The formulation.

21. 21. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHSGSTNYNPSLKS (SEQ ID NO: 47), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GX1X2NRPS (wherein X1 is N, Q, E or D and X2 is S or A) (SEQ ID NO: 20), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

22. 21. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: CDR-H1: GYYWS (SEQ ID NO: 16), CDR-H2: EINHAGSTNYNPSLKS (SEQ ID NO: 48), and CDR-H3: LPTRWVTTRYFDL (SEQ ID NO: 18); and a light chain variable domain comprising: CDR-L1: TGSSSNIGAGYDVH (SEQ ID NO: 19), CDR-L2: GDSNRPS (SEQ ID NO: 52), and CDR-L3: QSFDNSLSAYV (SEQ ID NO: 21).

23. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:

58.

24. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

7.

25. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:80; and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

7.

26. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable domain consisting of the amino acid sequence set forth in SEQ ID NO:

58.

27. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 2; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:

7.

28. The formulation of any one of claims 1 to 20, wherein the anti-ILT4 antibody or antigen-binding fragment thereof comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 80; and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:

7.

29. 29. The formulation of any one of claims 1 to 28, wherein the formulation is a liquid formulation stored at about 3°C ​​to about 5°C, frozen at or below -20°C, frozen at or below -70°C, or a reconstituted solution from a lyophilized formulation.

30. After 6 months of storage at about 3°C ​​to about 5°C: (i) the percent monomer of the anti-ILT4 antibody, or antigen-binding fragment thereof, is at least about 99% as measured by ultra-performance size exclusion chromatography; (ii) O.D. ３５０－５００ The turbidity of the formulation is up to about 0.135 when measured at (iii) the % main peak of the anti-ILT4 antibody or antigen-binding fragment thereof is at least about 63%, the % acidic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 23%, and / or the % basic variant of the anti-ILT4 antibody or antigen-binding fragment thereof is at most about 14%, as measured by high performance ion exchange chromatography; (iv) the number of sub-visible particles having a size of at least 2 μm is at most about 3500, as measured by microflow imaging; and / or (v) the percent oxidation of one or more amino acid residues selected from the group consisting of W7, W102, M253, M359, and M429 in the heavy chain of the anti-ILT4 antibody set forth in SEQ ID NO: 2 or 80 is less than about 4% as measured by reduced peptide mapping; A formulation according to any one of claims 1 to 27.

31. The formulation of any of claims 1 to 30, wherein the anti-ILT4 antibody or antigen-binding fragment thereof is a monoclonal antibody.

32. A method of treating cancer in a human patient in need thereof, comprising administering an effective amount of a formulation according to any one of claims 1 to 31.

33. 33. The method of claim 32, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma and small cell lung cancer.

34. Use of a formulation according to any one of claims 1 to 31 for the preparation of a medicament for treating cancer.

35. Use of a formulation according to any one of claims 1 to 31 for treating cancer in a human patient.

36. 35. The use according to claim 34, wherein the cancer is selected from the group consisting of colorectal cancer, esophageal cancer, melanoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma and small cell lung cancer.

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