Anti-IL-18BP antibody

Abstract

Antibodies that bind to interleukin-18 binding protein (IL-18BP), and antigen-binding fragments thereof, and related compositions are provided, which may be used in any of a variety of therapeutic and diagnostic methods, including the treatment or diagnosis of cancer and other diseases.

Description

[Background technology]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 437,526, filed January 6, 2023, and U.S. Provisional Patent Application No. 63 / 590,348, filed October 13, 2023, the contents of which are incorporated herein by reference in their entireties.

[0002] Interleukin-18 (IL-18) is an immunostimulatory cytokine with antitumor activity. This cytokine plays a key role in linking inflammatory immune responses with tumor progression. Indeed, recombinant human IL-18 has been evaluated as a cancer immunotherapy agent. However, this approach has not been successful, at least in part due to a human feedback loop in which administration of IL-18 increases the production of IL-18BP, which neutralizes the administered IL-18 (see, e.g., Robertson et al., Clinical Cancer Res. 12:4265-4273, 2006).

[0003] IL-18BP is a high-affinity IL-18 decoy receptor that is frequently upregulated in tumors. Research suggests that IL-18BP is a secreted immune checkpoint and a barrier to IL-18 immune activity (see, for example, Zhou et al., Nature. 583(7817):609-614, 2020). By sequestering IL-18 from its cell surface receptor, IL-18BP inhibits the pro-inflammatory activity of IL-18. The affinity of IL-18 for IL-18BP is higher than that of IL-18 for the IL-18 receptor, and IL-18BP is frequently present in amounts greater than IL-18, resulting in strict regulation. Furthermore, IL-18BP has been shown to balance immune responses, particularly Th1 and Th2, and plays an important role in autoimmune diseases (see, e.g., Park et al., Biomedicines. 10(7):1750, 2022). Thus, there is a need in the art for agents that specifically modulate the activity of IL-18BP. Summary of the Invention

[0004] The present disclosure relates to antibodies that bind to interleukin-18 binding protein (IL-18BP), and related compositions thereof, which may be used in any of a variety of therapeutic and diagnostic methods, including the treatment or diagnosis of cancer and other diseases.

[0005] Embodiments of the present disclosure include isolated antibodies, or antigen-binding fragments thereof, that bind to interleukin-18 binding protein (IL-18BP), wherein at least one antibody, or antigen-binding fragment thereof, comprises a V complementarity determining region selected from Table A1. H CDR1 sequence, V H CDR2 sequence, and V H A heavy chain variable region (V) containing CDR3 sequences and variants thereof that specifically bind to IL-18BP. H ), and a V of a complementarity-determining region selected from Table A1 L CDR1 sequence, V LCDR2 sequence, and V L A light chain variable region (V) containing CDR3 sequences and variants thereof that specifically bind to IL-18BP. L In certain embodiments, the compound includes: The VHCDR1, VHCDR2, and VHCDR3 sequences are TFX1X2X3X4X5H and IX6X7X8X9X, respectively. 10 X 11 X 12 X 13 X 14 X 15 AQKFQG, and X 16 X 17 X 18 X 19 X 20 X 21 X 22 DY, and the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each contain X 23 X 24 X 25 X 26 X 27 X 28 X 29 X 30 WX 31 A, X 32 X 33 X 34 X 35 X 36 X 37 X 38 , and QX 39 X 40 X 41 SFPYX 42 (See Table E11 for definition of "X" residues). V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 4 to 6, respectively; V H CDR1 sequence, V H CDR2 sequence, and V HThe CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 28 to 30, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 31 to 33, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 34 to 36, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 34 to 39, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 40 to 42, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 46 to 48, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 52 to 54, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1 sequence, V L CDR2 sequence, and VL the CDR3 sequences comprise SEQ ID NOs: 58 to 60, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 64 to 66, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 70 to 72, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 109 to 111, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 124 to 126, respectively; V HCDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 130 to 132, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 136 to 138, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 139 to 141, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 142 to 144, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 148 to 150, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 154 to 156, respectively; V H CDR1 sequence, V H CDR2 sequence, and V HThe CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 160 to 162, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 172 to 174, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1 sequence, VL CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 190 to 192, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 193 to 195, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 199 to 201, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 205 to 207, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 208-210, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 211 to 213, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 214-216, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 217 to 219, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 223 to 225, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 229 to 231, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 232 to 234, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 235 to 237, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 241 to 243, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 244 to 246, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 247 to 249, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 250 to 252, respectively; V HCDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 253 to 255, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 256 to 258, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 265 to 267, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 268-270, respectively; or V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 271 to 273, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences comprise SEQ ID NOs: 274 to 276, respectively.

[0006] In some embodiments, V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V H In some embodiments, V has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions. L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V L has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions. V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 286; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 288; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 292; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 294; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 296; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 298; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 352; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 353, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 354; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 355, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 356; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 357, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 358; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 359, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 360; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 361, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 362; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 368; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 369, and V L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:370.

[0007] Aspects of the present disclosure also include an isolated antibody or antigen-binding fragment thereof that binds to an epitope comprising I97 and V153 according to UniProt:O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO:1, or I67 and V123 as set forth in SEQ ID NO:2). In some embodiments, the isolated antibody or antigen-binding fragment thereof binds to interleukin-18 binding protein (IL-18BP) at an epitope comprising I97 and V153 according to UniProt:O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO:1, or I67 and V123 as set forth in SEQ ID NO:2).

[0008] In some embodiments: The VHCDR1, VHCDR2, and VHCDR3 sequences are TFX1X2X3X4X5H and IX6X7X8X9X, respectively. 10 X 11 X 12 X 13 X 14 X 15 AQKFQG, and X 16 X 17 X 18 X 19 X 20 X 21 X 22 DY, and the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each contain X 23 X 24 X 25 X 26 X 27 X 28 X 29 X 30 WX 31 A, X 32 X 33 X 34 X 35 X 36 X 37 X 38 , and QX 39 X 40 X 41 SFPYX 42 (See Table E11 for definition of "X" residues). V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 4-6, respectively; or V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 265 to 267, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences comprise SEQ ID NOs: 268 to 270, respectively.

[0009] In certain embodiments, V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and V L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:368.

[0010] Aspects of the present disclosure also include an isolated antibody or antigen-binding fragment thereof, wherein the antibody binds to interleukin-18 binding protein (IL-18BP) and competes with IL-18 for binding of IL-18BP. In another aspect, the present disclosure provides an antibody that binds to IL-18BP and interferes with the binding of IL-18BP to IL-18. In another aspect, the present disclosure provides an antibody that binds to IL-18BP and is an antagonist of IL-18BP, wherein the antibody antagonizes the binding activity between IL-18BP and IL-18. In some embodiments, the present disclosure provides an antibody that binds to a preformed IL-18-IL-18BP complex. In some embodiments, the present disclosure provides an antibody that binds to free IL-18BP. In some embodiments, the antibody or antigen-binding fragment thereof binds to a conformational epitope of IL-18BP. In some embodiments, the conformational epitope comprises two or more amino acid residues selected from the group consisting of K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO: 372. In some embodiments, the conformational epitope comprises amino acid residues K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO: 372. In some embodiments, the antibody or antigen-binding fragment thereof binds to a linear epitope of IL-18BP. In some embodiments, the isolated antibody or antigen-binding fragment thereof binds to the binding interface between IL-18 and mature IL-18BP. In some embodiments, the isolated antibody or antigen-binding fragment thereof binds to amino acid residues R61, Y69, and R131 of SEQ ID NO: 372. In some embodiments, the isolated antibody or antigen-binding fragment thereof comprises a VH comprising a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and a VL comprising a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334.

[0011] In some embodiments, the isolated antibody or antigen-binding fragment thereof binds to human IL-18BP and cynomolgus IL-18BP, but does not bind (specifically or substantially) to mouse IL-18BP. In some embodiments, the isolated antibody or antigen-binding fragment thereof binds to human IL-18BP, cynomolgus IL-18BP, and mouse IL-18BP. In some embodiments, the isolated antibody or antigen-binding fragment thereof binds to the binding interface between IL-18 and mature IL-18BP. In certain embodiments, the isolated antibody or antigen-binding fragment thereof binds to the binding affinity (K D The isolated antibody or antigen-binding fragment thereof binds to IL-18BP with a binding affinity of greater than about 650 pM, and optionally with a binding affinity of between about 1 pm and about 650 pm, or about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM or less. In some embodiments, the isolated antibody or antigen-binding fragment thereof is an IL-18BP antagonist that antagonizes the binding activity between IL-18BP and IL-18. In some embodiments, the isolated antibody or antigen-binding fragment thereof blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signaling, including induction of IFN-gamma, CXCL10 and / or TNFα.

[0012] In some embodiments, the isolated antibody or antigen-binding fragment thereof comprises an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or a hybrid and / or variant thereof. In some embodiments, the isolated antibody or antigen-binding fragment thereof comprises an IgG Fc domain with high effector function in humans, optionally an IgG1 or IgG3 Fc domain. In some embodiments, the isolated antibody or antigen-binding fragment thereof comprises an IgG Fc domain with reduced effector function in humans, optionally an IgG2 or IgG4 Fc domain. In some embodiments, the isolated antibody or antigen-binding fragment thereof is a monoclonal antibody. In some embodiments, the isolated antibody or antigen-binding fragment thereof is a humanized antibody. In some embodiments, the isolated antibody or antigen-binding fragment thereof is selected from an Fv fragment, a single-chain Fv (scFv) polypeptide, an adnectin, anticalin, an aptamer, an avimer, a camelid antibody, a designed ankyrin repeat protein (DARPin), a minibody, a nanobody, and a unibody.

[0013] Also included are isolated polynucleotides encoding the isolated anti-IL-18BP antibodies, or antigen-binding fragments thereof, described herein, expression vectors containing the isolated polynucleotides, and isolated host cells containing the vectors. Also provided are one or more isolated polynucleotides encoding the anti-IL-18BP antibodies described herein. For example, the V of the antibodies disclosed herein H A first polynucleotide encoding a region and a V region of the antibody disclosed herein L A second polynucleotide encoding the region is provided herein.

[0014] Certain embodiments include pharmaceutical compositions comprising an isolated anti-IL-18BP antibody, or antigen-binding fragment thereof, described herein and a pharmaceutically acceptable carrier. In some embodiments, the composition has a purity of at least about 80%, 85%, 90%, 95%, 98%, or 99% on a protein basis with respect to at least one antibody or antigen-binding fragment, and is substantially free of aggregates and endotoxins. In some embodiments, the composition is optionally a sterile injectable solution suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

[0015] Also included are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some embodiments, the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, optionally a proliferative disease or disorder selected from lymphoproliferative disorders, myeloproliferative disorders, proliferative enterocolitis, proliferative diabetic retinopathy, and proliferative renal diseases. In some embodiments, the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18. In some embodiments, the cancer is selected from the group consisting of bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (liver cell carcinoma), and / or hepatocellular carcinoma (liver cell carcinoma). and / or ovarian cancer), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, urothelial cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer.

[0016] Certain embodiments include administering the pharmaceutical composition (comprising an anti-IL18BP antibody or antigen-binding fragment thereof) in combination with IL-18. Some embodiments include administering the pharmaceutical composition (comprising an anti-IL18BP antibody or antigen-binding fragment thereof) in combination with an immune checkpoint modulator selected from an antagonist of an inhibitory immune checkpoint molecule and an agonist of a stimulatory immune checkpoint molecule. In some embodiments, the immune checkpoint modulator is a polypeptide, optionally an antibody or antigen-binding fragment thereof, or a ligand, or a small molecule. In some embodiments, the inhibitory immune checkpoint molecule is selected from one or more of Programmed Death-Ligand 1 (PD-L1), Programmed Death 1 (PD-1), Programmed Death-Ligand 2 (PD-L2), Cytotoxic T-Lymphocyte-Associated protein 4 (CTLA-4), Indoleamine 2,3-dioxygenase (IDO), Tryptophan 2,3-dioxygenase (TDO), T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3), Lymphocyte Activation Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T Lymphocyte Attenuator (BTLA), CD160, Herpes Virus Entry Mediator (HVEM), and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0017] In some embodiments: the antagonist is an antibody or antigen-binding fragment or small molecule that specifically binds to PD-L1 and / or PD-L2, an antagonist of PD-L1 and / or PD-L2 optionally chosen from one or more of atezolizumab (MPDL3280A), avelumab (MSB0010718C) and durvalumab (MEDI4736), optionally in which the cancer is selected from one or more of colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer and renal cell carcinoma; the antagonist is a PD-1 antagonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally in which the PD-1 antagonist is nivolumab, and the cancer is optionally chosen from one or more of Hodgkin lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer; the PD-1 antagonist is pembrolizumab, and the cancer is optionally selected from one or more of melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial cancer; the antagonist is a CTLA-4 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, optionally in which the cancer is selected from melanoma, prostate cancer, lung cancer, and bladder cancer; the antagonist is an IDO antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-l-tryptophan (1MT), β-carboline (norharman; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, wherein the cancer is optionally selected from one or more of metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or malignant brain tumor; the antagonist is a TDO antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to TDO, 680C91 and LM10; The antagonist is a TIM-3 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to TIM-3; the antagonist is a LAG-3 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to LAG-3, and BMS-986016; The antagonist is a VISTA antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to VISTA; the antagonist is an antagonist of BTLA, CD160 and / or HVEM optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to BTLA, CD160 and / or HVEM; The antagonist is a TIGIT antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to TIGIT.

[0018] In some embodiments, the stimulatory immune checkpoint molecule is selected from one or more of OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

[0019] In certain embodiments: the agonist is an OX40 agonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L, and GSK3174998; the agonist is a CD40 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD40, CP-870,893, dacetuzumab, Chi Lob 7 / 4, ADC-1013, and rhCD40L, wherein the cancer is optionally selected from one or more of melanoma, pancreatic cancer, mesothelioma, and hematological cancers, optionally lymphomas such as non-Hodgkin's lymphoma; the agonist is a GITR agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to GITR, INCAGN01876, DTA-1, and MEDI1873; the agonist is a CD137 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD137, utomilumab, and 4-1BB ligand; the agonist is a CD27 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD27, varlilumab, and CDX-1127 (1F5); the agonist is a CD28 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD28, and TAB08; and / or The agonist is an HVEM agonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule or a ligand that specifically binds to HVEM.

[0020] Certain embodiments include administering a pharmaceutical composition (comprising an anti-IL18BP antibody or antigen-binding fragment thereof) in combination with at least one chemotherapeutic agent, in some embodiments, the at least one chemotherapeutic agent is selected from one or more of an alkylating agent, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor (type 1 or type II), and an anti-microtubule agent.

[0021] In some embodiments: the alkylating agent is selected from one or more of nitrogen mustards (optionally mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosoureas (optionally N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, and streptozotocin), tetrazines (optionally dacarbazine, mitozolomide, and temozolomide), aziridines (optionally thiotepa, mitomycin, and diaziquone (AZQ)), cisplatin and its derivatives (optionally carboplatin and oxaplatin), and non-classical alkylating agents (optionally procarbazine and hexamethylmelamine); the antimetabolite is selected from one or more of antifolates (optionally methotrexate and pemetrexed), fluoropyrimidines (optionally 5-fluorouracil and capecitabine), deoxynucleoside analogs (optionally ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (optionally thioguanine and mercaptopurine); the cytotoxic antibiotic is selected from one or more of anthracyclines (optionally doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, and mitoxantrone), bleomycin, mitomycin C, mitoxantrone, and actinomycin; the topoisomerase inhibitor is selected from one or more of camptothecin, irinotecan, topotecan, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, mervalone, and aclarubicin; and / or The anti-microtubule agent is selected from one or more of taxanes (optionally paclitaxel and docetaxel) and vinca alkaloids (optionally vinblastine, vincristine, vindesine, vinorelbine).

[0022] In some embodiments, the disease or condition is myelodysplastic syndrome (MDS). In some embodiments, the disease or condition is an infectious disease. In certain embodiments, the infectious disease is selected from viral, bacterial, fungal (optionally yeast), and protozoal infections. Certain embodiments include administering a pharmaceutical composition (comprising an anti-IL18BP antibody or antigen-binding fragment thereof) in combination with IL-18.

[0023] Also provided is a method of screening an anti-IL-18BP antibody or antigen-binding fragment thereof for the ability to block or inhibit binding between IL-18 and IL-18BP, the method comprising: (a) determining the binding affinity of an antibody or antigen-binding fragment thereof to (i) IL-18BP alone, and (ii) an IL-18-reduced fusion protein, the IL-18-reduced fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and an optional protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein is linked to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein; (b) comparing the binding affinity of (i) with the binding affinity of (ii); and (c) identifying or selecting an antibody or antigen-binding fragment thereof as being capable of blocking or inhibiting the binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii).

[0024] In some embodiments, the IL-18 and IL-18BP are murine IL-18 and IL-18BP. In some embodiments, the IL-18 and IL-18BP are human IL-18 and IL-18BP. In some embodiments, the low IL-18 fusion protein comprises, from N- to C-terminus, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP. In some embodiments, the low IL-18 fusion protein comprises an amino acid sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

[0025] Certain embodiments include an isolated low IL-18 fusion protein comprising, from N- to C-terminus, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP. In some embodiments, the low IL-18 fusion protein comprises, consists of, or consists essentially of an amino acid sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

[0026] Also included are methods of stimulating an immune response in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some embodiments, the immune response is an IL-18-mediated immune response. In certain embodiments, the IL-18-mediated immune response comprises induction of IFN-gamma, CXCL10, and / or TNFα in a subject in need thereof. [Brief explanation of the drawings]

[0027] [Figure 1] FIG. 1 shows the family of antibodies of the present disclosure. [Figure 2]Figure 2 shows an alignment of cynomolgus (SEQ ID NO: 373), human (SEQ ID NO: 371), and mouse (SEQ ID NO: 375) IL-18BP orthologs. The signal peptide (absent in the mature peptide) is underlined. Four regions marked with downward arrows (↓) indicate amino acids that are identical in human and mouse but differ in cynomolgus IL-18BP. [Figure 3A] Figure 3A shows the design of the low IL-18. Figure 3A shows a schematic of the low IL-18 expression cassette. The genetic landmarks from the N' to C' terminus include the osteonectin signal peptide, the human IL-18 coding region, a flexible gly-ser linker interrupted by a tobacco etch virus (TEV) protease cleavage site, the human IL-18BP coding sequence, and a 6x HIS tag. [Figure 3B] Figure 3B shows the design of a low IL-18. Figure 3B shows an outline of a model derived from the crystal structure of human IL-18 complexed with ectromelia virus IL-18BP (shown). The N-terminus of IL-18BP and the C-terminus of IL-18 are indicated by boxes and arrows. [Figure 4] Figure 4 shows the suppression of IFNγ expression by anti-IL-18BP mAb in human KG-1 cells. mAb1191 (R&D Systems) is a commercially available mouse anti-human IL-18BP neutralizing antibody and was used as a positive control in this experiment (catalog number mab1191). [Figure 5] FIG. 5 shows anti-mIL-18BP mAbs tested in a mouse IL-18 reporter assay. [Figure 6] Figure 6 shows anti-mIL-18BP mAbs tested in a mouse splenocyte assay. There was no clear upper plateau, so IC50 values ​​could not be determined. [Figure 7A]Figure 7A shows the antitumor activity of mIL-18 in combination with SA51d (SA0051d) in established MC38 colon tumors. Figure 7A shows tumor growth of MC38 cells subcutaneously implanted into C57BL / 6 mice to form tumors. Fifty mice were then randomized into five groups (n = 10) and treated with either PBS vehicle control, mIL-18, SA51d, or the combination of mIL-18 and SA51d. Animals were administered up to seven total doses every three days, as indicated by arrows. The average tumor size for each group was calculated and graphed. The combination of mIL-18 and SA0051d was able to significantly inhibit growth compared to the control. [Figure 7B] Figure 7B shows the antitumor activity of mIL-18 in combination with SA51d (SA0051d) in established MC38 colon tumors. Figure 7B shows that the combination of mIL-18 + SA51d was able to significantly inhibit growth by day 15 compared to the vehicle control (p<0.0001). [Figure 8A] Figure 8A shows the evaluation of mAbs in an IL-18-mediated HEK293 reporter assay. Figure 8A shows pre-incubation of mAbs with IL-18BP. [Figure 8B] Figure 8B shows evaluation of mAbs in an IL-18-mediated HEK293 reporter assay. Figure 8B shows pre-incubation of IL-18 with IL-18BP. [Figure 9] Figure 9 shows the effect of mAbs on IFNγ secretion in KG-1 cells. Complex alone represents the addition of IL-18BP and mAb in the absence of IL-18. [Figure 10A] Figure 10A shows induction of IFNγ responses in human (10A) PBMCs by high-affinity mAbs. Dose response of IFNγ production in response to mAb inhibition of IL-18BP. 2 x 10 PBMCs / well were incubated with 1 ng / mL IL-12 and 2 ng / mL IL-18, along with increasing concentrations of each mAb. Supernatant aliquots were harvested at 48 hours and assayed for IFNγ by ELISA. [Figure 10B]10B shows induction of IFNγ responses in cynomolgus monkey (10B) PBMCs by high affinity mAbs. Dose response of IFNγ production in response to mAb inhibition of IL-18BP. 2×105 PBMCs / well were incubated with 1 ng / mL IL-12 and 2 ng / mL IL-18, along with increasing concentrations of each mAb. Supernatant aliquots were harvested at 48 hours and assayed for IFNγ by ELISA. [Figure 11A] FIG. 11A shows the induction of IFNγ responses in PBMC from pre-complexed IL-18 / IL-18BP by high affinity mAbs. [Figure 11B] FIG. 11B shows the induction of IFNγ responses in PBMCs from pre-complexed IL-18 / IL-18BP by high affinity mAbs. [Figure 12] Figure 12 shows a co-crystal structure model of the IL-18 / IL-18BP complex. The locations of four differences in IL-18BP and IL-18 shared by humans and mice, but not cynomolgus monkeys, are shown in dark font (V75M, I97M, R113Q, V153M) and light font (V47I, T99A, K115R, F170Y), respectively. Note that I98M and V153M are close and present at the binding interface, while V75M and R113Q are not near the active site. [Figure 13] Figure 13 shows mAbs binding to human IL-18BP with two orthologous cynomolgus amino acid substitutions. The measured kinetic quality is summarized in the associated table as "-" for no binding and 1 to 4+ for varying degrees of binding. [Figure 14] Figure 14 shows the conformational epitope of SA64a determined by XL-MS mass spectrometry. A partial sequence of IL-18BP is shown with the residues cross-linked to SA64a designated as the epitope. [Figure 15-1]Figure 15 shows amino acid variants of the SA01a antibody from combinatorial screening that retain or improve binding to IL-18BP (x = good binding for humans, cynomolgus monkeys, and mice; hc = good binding for humans and cynomolgus monkeys, but not mice; hm = good binding for humans and mice, but not cynomolgus monkeys; cm = good binding for cynomolgus monkeys and humans, but not humans; m = good binding for mice, but not humans and cynomolgus monkeys). [Figure 15-2] Figure 15 shows amino acid variants of the SA01a antibody from combinatorial screening that retain or improve binding to IL-18BP (x = good binding for humans, cynomolgus monkeys, and mice; hc = good binding for humans and cynomolgus monkeys, but not mice; hm = good binding for humans and mice, but not cynomolgus monkeys; cm = good binding for cynomolgus monkeys and humans, but not humans; m = good binding for mice, but not humans and cynomolgus monkeys). [Figure 16] Figure 16 shows spider plots of the efficacy of anti-mouse IL-18BP antibodies in the MC38 syngeneic tumor model. The results demonstrate significantly improved efficacy using anti-mouse PD-1 + anti-mouse IL-18BP (SA51d, or SA0051d) + mIL-18 compared with anti-mouse PD-1 alone or anti-mouse PD-1 + mIL-18. [Figure 17] Figure 17 shows levels of IL-18 (left) and IFNγ (right) in MC38 tumors from anti-IL-18BP-treated mice compared to vehicle-treated animals. Tumors were harvested at the endpoint of the efficacy study and pro-inflammatory cytokines were assessed by ELISA. [Figure 18]Figure 18 shows relative NK cell marker (left) and granzyme B (right) expression in MC38 tumors harvested at efficacy study endpoint, as assessed by qPCR. Markers of NK cell numbers and activity increased in response to anti-IL-18BP and further increased with anti-PD-1 combination therapy. [Figure 19A-1] Figure 19A shows the efficacy of anti-mouse IL-18BP antibodies in the MC38 syngeneic tumor re-challenge model. Results showed sustained responses in 11 of 12 animals upon re-challenge. [Figure 19A-2] Figure 19A shows the efficacy of anti-mouse IL-18BP antibodies in the MC38 syngeneic tumor re-challenge model. Results showed sustained responses in 11 of 12 animals upon re-challenge. [Figure 19B-1] Figure 19B shows the efficacy of anti-mouse IL-18BP antibodies in the MC38 syngeneic tumor re-challenge model. Results showed sustained responses in 11 of 12 animals upon re-challenge. [Figure 19B-2] Figure 19B shows the efficacy of anti-mouse IL-18BP antibodies in the MC38 syngeneic tumor re-challenge model. Results showed sustained responses in 11 of 12 animals upon re-challenge. [Figure 19B-3] Figure 19B shows the efficacy of anti-mouse IL-18BP antibodies in the MC38 syngeneic tumor re-challenge model. Results showed sustained responses in 11 of 12 animals upon re-challenge. [Figure 20] Figure 20 shows the production of CXCL10 (left) and CCL2 (right) in human PBMCs in response to high-affinity mAbs. CXCL10 and CCL2 were secreted with the addition of IL-12 and IL-18, as assessed by ELISA, and were enhanced by the addition of mAbs. [Figure 21A]Figure 21A shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in an EMT6 mouse syngeneic tumor model. Figure 21A shows that survival of EMT6 tumor-bearing mice treated with the anti-IL-18BP antibody SA0051d was significantly enhanced, and was further improved by combination with an anti-mouse PD-1 antibody. Treatment with SA0051d reduced tumor growth, and tumor regression was observed in some animals in combination with anti-PD-1. [Figure 21B] Figure 21B shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in an EMT6 mouse syngeneic tumor model. Figure 21B shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, and tumor regression was observed in some animals in combination with anti-PD-1. [Figure 21C] Figure 21C shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in an EMT6 mouse syngeneic tumor model. Figure 21C shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, and tumor regression was observed in some animals in combination with anti-PD-1. [Figure 21D] Figure 21D shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in an EMT6 mouse syngeneic tumor model. Figure 21D shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, and in combination with anti-PD-1, tumor regression was observed in some animals. [Figure 21E] Figure 21E shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in an EMT6 mouse syngeneic tumor model. Figure 21B shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, and tumor regression was observed in some animals in combination with anti-PD-1. [Figure 22A]Figure 22A shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in the E0771 mouse syngeneic tumor model. Figure 22A shows the mean tumor growth for each group. Treatment with SA0051d reduced tumor growth, as did treatment with the combination of SA0051d and an anti-mouse PD-1 antibody. [Figure 22B] Figure 22B shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in the E0771 mouse syngeneic tumor model. Figure 22B shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, as did treatment with the combination of SA0051d and an anti-mouse PD-1 antibody. [Figure 22C] Figure 22C shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in the E0771 mouse syngeneic tumor model. Figure 22C shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, as did treatment with a combination of SA0051d and an anti-mouse PD-1 antibody. [Figure 22D] Figure 22D shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in the E0771 mouse syngeneic tumor model. Figure 22D shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, as did treatment with a combination of SA0051d and an anti-mouse PD-1 antibody. [Figure 22E] Figure 22E shows the efficacy of the anti-mouse IL-18BP antibody SA0051d in the E0771 mouse syngeneic tumor model. Figure 22E shows spider plots monitoring tumor growth in each animal. Treatment with SA0051d reduced tumor growth, as did treatment with a combination of SA0051d and an anti-mouse PD-1 antibody. DETAILED DESCRIPTION OF THE INVENTION

[0028] The present disclosure relates to antibodies and antigen-binding fragments thereof that specifically bind to interleukin-18 binding protein (IL-18BP), e.g., human IL-18BP, particularly antibodies and antigen-binding fragments thereof with epitope specificity and improved properties. Some embodiments include certain humanized antibodies and fragments thereof that can bind to IL-18BP and block or reduce the inhibitory binding of IL-18BP to its ligand, IL-18, thereby increasing IL-18-mediated downstream signaling. In certain embodiments, the anti-IL-18BP antibody or antigen-binding fragment thereof is an antagonist or inhibitor of IL-18BP.

[0029] The IL-18BP antagonist antibodies described herein are useful for the treatment and prevention of various diseases and conditions, such as cancer. Accordingly, some embodiments relate to the use of anti-IL-18BP antibodies, or antigen-binding fragments thereof, for the diagnosis, evaluation, and treatment of diseases and conditions, including those associated with IL-18 and / or those associated with the activity or aberrant expression of IL-18BP.

[0030] The practice of the present disclosure will employ, unless specifically indicated to the contrary, conventional methods of virology, immunology, microbiology, molecular biology, and recombinant DNA technology within the skill of the art, many of which are described below for illustrative purposes. Such techniques are fully explained in the literature. See, for example, Current Protocols in Molecular Biology or Current Protocols in Immunology, John Wiley & Sons, New York, NY (2009); Ausubel et al., Short Protocols in Molecular Biology, 3 rded., Wiley & Sons, 1995; Sambrook and Russell, Molecular Cloning: A Laboratory Manual (3rd Edition, 2001); Maniatis et al. Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., 1984); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., 1985); Transcription and Translation (B. Hames & S. Higgins, eds., 1984); Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984), and other similar references.

[0031] definition As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the content clearly dictates otherwise.

[0032] By "about" is meant an amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% relative to the reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length.

[0033] The term "antigen" refers to a molecule or portion of a molecule that can be bound by a selective binding agent, such as an antibody, and can be used in an animal to generate antibodies capable of binding to an epitope of that antigen. An antigen may have one or more epitopes. As used herein, the term "antigen" includes a substance that, under appropriate conditions, is capable of inducing an immune response against the substance and reacting with the products of the immune response. For example, an antigen can be recognized by antibodies (humoral immune response) or sensitized T lymphocytes (T helper or cell-mediated immune response), or both. Antigens can be soluble substances, such as toxins and foreign proteins, or microparticles, such as bacteria and tissue cells; however, only portions of a protein or polysaccharide molecule, known as an antigenic determinant (epitope), combine with antibodies or specific receptors on lymphocytes. More broadly, the term "antigen" includes any substance to which an antibody binds or for which an antibody is desired, regardless of whether the substance is immunogenic. For such antigens, antibodies can be identified by recombinant methods, independent of any immune response.

[0034] "Antagonist" refers to an agent (e.g., an antibody) that interferes with or otherwise reduces the physiological action of another agent or molecule. In some instances, antagonists specifically bind to other agents or molecules. Included are full and partial antagonists.

[0035] "Agonist" refers to an agent (e.g., an antibody) that increases or enhances the physiological effect of another agent or molecule. In some instances, agonists specifically bind to other agents or molecules. Included are full and partial agonists.

[0036] As used herein, the term "amino acid" is intended to refer to both natural and unnatural amino acids, as well as amino acid analogs and mimetics. Natural amino acids include the 20 (L) amino acids utilized during protein biosynthesis, as well as others, such as 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, homocysteine, citrulline, and ornithine. Unnatural amino acids include, for example, (D)-amino acids, norleucine, norvaline, p-fluorophenylalanine, ethionine, and the like, which are known to those skilled in the art. Amino acid analogs include modified forms of natural and unnatural amino acids. Such modifications can include, for example, the substitution or exchange of chemical groups and moieties on the amino acid or by derivatization of the amino acid. Amino acid mimetics include, for example, organic structures that exhibit functionally similar properties, such as the charge and charge-spacing characteristics of the reference amino acid. For example, an organic structure that mimics arginine (Arg or R) will have a positively charged portion positioned in a similar molecular space and with the same degree of flexibility as the e-amino group of the side chain of the natural Arg amino acid. Mimetics also include structures that are constrained to maintain optimal spacing and charge interactions of amino acids or amino acid functional groups. Those skilled in the art will know or be able to determine which structures constitute functionally equivalent amino acid analogs and amino acid mimetics.

[0037] As used herein, the term "antibody" includes not only intact polyclonal or monoclonal antibodies, but also fragments thereof (e.g., dAb, Fab, Fab', F(ab'), Fv, etc.), single chain (scFv), synthetic variants thereof, naturally occurring variants, fusion proteins comprising an antibody portion with an antigen-binding fragment of the required specificity, humanized antibodies, chimeric antibodies, and any other modified configuration of an immunoglobulin molecule that contains an antigen-binding site or fragment (epitope-recognition site) of the required specificity. Specific features and characteristics of antibodies (and antigen-binding fragments thereof) are described in more detail herein.

[0038] The antibody or antigen-binding fragment can be of essentially any type. As is well known in the art, an antibody is an immunoglobulin molecule capable of specifically binding to a target, such as an immune checkpoint molecule, through at least one epitope recognition site located in the variable region of the immunoglobulin molecule.

[0039] As used herein, the term "antigen-binding fragment" refers to a polypeptide fragment that contains at least one CDR of an immunoglobulin heavy chain and / or light chain that binds to an antigen of interest. In this regard, an antigen-binding fragment of an antibody described herein is a V CDR from an antibody that binds to a target molecule. H and V L In certain embodiments, the antigen-binding fragments of the present disclosure may comprise one, two, three, four, five, or all six CDRs of the V of an antibody disclosed herein. H Array and V L The sequence includes all six CDRs.

[0040] The binding properties of antibodies and antigen-binding fragments thereof can be quantified using methods well known in the art (see Davies et al., Annual Rev. Biochem. 59:439-473, 1990). In some embodiments, the antibody or antigen-binding fragment thereof binds to a target molecule, e.g., an IL-18BP polypeptide or epitope or complex thereof, with a binding affinity of about ≦10 -7 M ~ about 10 -8 M. In some embodiments, the equilibrium dissociation constant is about ≦10 -9 M~approx.≦10 -10In certain exemplary embodiments, the antibody or antigen-binding fragment thereof has an affinity (K) for the target molecule (to which it specifically binds) of less than about, at least about, or about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, or 50 nM. D or EC 50 )

[0041] A molecule, such as a polypeptide or antibody, is said to exhibit "specific binding" or "preferential binding" if it reacts or associates with a particular cell, substance, or particular epitope more frequently, more rapidly, with greater duration, and / or with greater affinity than it does with alternative cells, substances, or epitopes. An antibody "specifically binds" or "preferentially binds" to a target molecule or epitope if it binds, for example, by a statistically significant amount, with greater affinity, avidity, more readily, and / or with longer duration than it binds to other substances or epitopes. Typically, one member of a pair of molecules exhibiting specific binding has a region or cavity on its surface that specifically binds to, and is therefore complementary to, a particular spatial and / or polar structure of the other member of the pair of molecules. In this way, the members of the pair have the property of specifically binding to each other. For example, an antibody that specifically or preferentially binds to a specific epitope is one that binds to that specific epitope with greater affinity, avidity, more readily, and / or with longer duration than it binds to other epitopes. It is also understood by reading this definition that, for example, an antibody (or moiety or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. This term is also applicable, for example, when an antibody is specific for a particular epitope carried by multiple antigens, in which case a specific binding member carrying an antigen-binding fragment or domain can bind to various antigens that carry the epitope; for example, it may be cross-reactive with multiple different forms of a target antigen from multiple species that share a common epitope.

[0042] Immunological binding generally refers to the type of non-covalent interaction that occurs between an immunoglobulin molecule and an antigen for which the immunoglobulin is specific, for example, by way of illustration and not limitation, as a result of electrostatic, ionic, hydrophilic and / or hydrophobic attractions or repulsions, steric forces, hydrogen bonding, van der Waals forces, and other interactions. The strength, or affinity, of an immunological binding interaction is determined by the dissociation constant (K D ) and smaller K D represents a greater affinity. The immunological binding properties of a selected polypeptide can be quantified using methods well known in the art. One such method involves measuring the rates of antigen-binding site / antigen complex formation and dissociation, where these rates depend on the concentrations of the complex partners, the affinity of the interaction, and geometric parameters that affect the rates equally in both directions. In this way, both the "on-rate constant" (Kon) and the "dissociation rate constant" (Koff) can be determined by calculation of the concentrations and the actual rates of association and dissociation. The ratio of Koff / Kon allows for the removal of all parameters not related to affinity, and thus the dissociation constant K D As used herein, the term "affinity" includes the equilibrium constant for the reversible binding of two drugs, K D or EC 50 The affinity of a binding protein to a ligand, such as the affinity of an antibody for an epitope, can be, for example, about 100 nanomolar (nM) to about 0.1 nM, about 100 nM to about 1 picomolar (pM), or about 100 nM to about 1 femtomolar (fM). As used herein, the term "avidity" refers to the resistance of a complex of two or more agents to dissociation after dilution. In some embodiments, affinity is measured using the half-maximal effective concentration (EC 50 ) and refers to the concentration of an agent, such as an antibody or anti-IL-18BP antibody, as disclosed herein, that induces a response midway between baseline and maximum after a specified exposure time. 50is commonly used as a measure of antibody potency.

[0043] Antibodies can be prepared by any of a variety of techniques known to those of skill in the art. See, e.g., Harlow and Lane, *Antibodies: A Laboratory Manual*, Cold Spring Harbor Laboratory, 1988. Monoclonal antibodies specific for a polypeptide of interest can be prepared, for example, using the technique of Kohler and Milstein, *Eur. J. Immunol.* 6:511-519, 1976, and modifications thereto. Also included are methods for expressing human antibodies using transgenic animals, such as mice. See, e.g., Neuberger et al., *Nature Biotechnology* 14:826, 1996; Lonberg et al., *Handbook of Experimental Pharmacology* 113:49-101, 1994; and Lonberg et al., *Internal Review of Immunology* 13:65-93, 1995. A particular example includes the VELOCIMMUNE® platform by REGENEREX® (see, eg, US Pat. No. 6,596,541).

[0044] Antibodies can also be generated or identified using phage display libraries or yeast display libraries (see, e.g., U.S. Patent No. 7,244,592; Chao et al., Nature Protocols. 1:755-768, 2006). Non-limiting examples of available libraries include cloned or synthetic libraries, such as the human combinatorial antibody library (HuCAL), in which the structural diversity of the human antibody repertoire is represented by seven heavy chain and seven light chain variable region genes. Combinations of these genes generate 49 frameworks in the master library. By stacking highly variable gene cassettes (CDRs = complementarity-determining regions) on these frameworks, a vast human antibody repertoire can be reproduced. Also included are human libraries designed using human donor-supplied fragments encoding the light chain variable region, synthetic DNA encoding the heavy chain CDR-3, heavy chain CDR-1 diversity, and synthetic DNA encoding the heavy chain CDR-2 diversity. Other libraries suitable for use will be apparent to those skilled in the art.

[0045] In certain embodiments, the antibodies and antigen-binding fragments thereof described herein comprise a set of heavy and light chain CDRs, each inserted between a set of heavy and light chain framework regions (FRs), which provide support for the CDRs and define the spatial relationship of the CDRs to each other. As used herein, the term "CDR set" refers to the three hypervariable regions of a heavy or light chain V region. Proceeding from the N-terminus of the heavy or light chain, these regions are designated "CDR1," "CDR2," and "CDR3," respectively. An antigen-binding site therefore comprises six CDRs, including a set of CDRs from each of the heavy and light chain V regions. A polypeptide comprising a single CDR (e.g., CDR1, CDR2, or CDR3) is referred to herein as a "molecular recognition unit." Crystallographic analysis of numerous antigen-antibody complexes has demonstrated that the amino acid residues of the CDRs form extensive contacts with the bound antigen, with the most extensive antigen contacts being with the heavy chain CDR3. Thus, the molecular recognition unit is primarily responsible for the specificity of the antigen-binding site.

[0046] As used herein, the term "FR set" refers to four adjacent amino acid sequences that frame the CDRs of a CDR set of a heavy or light chain V region. Some FR residues may contact the bound antigen; however, FRs are primarily involved in folding the V region into the antigen-binding site, particularly the FR residues directly adjacent to the CDRs. Within FRs, certain amino acid residues and certain structural features are highly conserved. In this regard, most V region sequences contain an internal disulfide loop of approximately 90 amino acid residues. When the V region folds into the binding site, the CDRs are presented as protruding loop motifs that form the antigen-binding surface. Regardless of the exact CDR amino acid sequence, it is generally recognized that conserved structural regions of FRs that affect the folded shape of the CDR loops exist in certain "canonical" structures. Furthermore, certain FR residues are known to participate in non-covalent interdomain contacts that stabilize the interaction of antibody heavy and light chains.

[0047] The structure and location of immunoglobulin variable domains can be determined by reference to Kabat, EA et al., Sequences of Proteins of Immunological Interest. 4th Edition. U.S. Department of Health and Human Services, 1987, and updates thereof.

[0048] Also included are monoclonal antibodies, which refer to homogeneous antibody populations composed of amino acids (natural and non-natural) involved in selective binding of an epitope. The term "monoclonal antibody" encompasses not only intact and full-length monoclonal antibodies, but also fragments thereof (e.g., Fab, Fab', F(ab')2, Fv, etc.), single chain (scFv), variants thereof, fusion proteins containing the antigen-binding portion, humanized monoclonal antibodies, chimeric monoclonal antibodies, and any other modified configuration of an immunoglobulin molecule containing an antigen-binding fragment (epitope recognition site) of the required specificity and ability to bind to the epitope. It is not intended to be limited with regard to the source of the antibody or the manner in which it is made (e.g., by hybridoma, phage selection, recombinant expression, transgenic animals). The term includes whole immunoglobulins, as well as fragments such as those described above under the definition of "antibody."

[0049] The proteolytic enzyme papain preferentially cleaves IgG molecules to produce several fragments, two of which (F(ab) fragments) each comprise a covalently linked heterodimer containing an intact antigen-binding site. The enzyme pepsin can cleave IgG molecules to provide several fragments, including the F(ab')2 fragment, which contains both antigen-binding sites. Fv fragments for use according to certain embodiments can be produced by preferential proteolytic cleavage of IgM, and in rare cases, IgG or IgA immunoglobulin molecules. Fv fragments, however, are more commonly derived using recombinant techniques known in the art. Fv fragments comprise a noncovalent VH::VL heterodimer containing an antigen-binding site that retains much of the antigen recognition and binding capabilities of a native antibody molecule (Inbar et al., PNAS USA. 69:2659-2662, 1972; Hochman et al., Biochem. 15:2706-2710, 1976; and Ehrlich et al., Biochem. 19:4091-4096, 1980). In some embodiments, the Fv is stabilized by other means, such as the incorporation of at least one disulfide bond (Worn & Pluckthun, J. Mol. Biol. 305, 989-1010, 2001).

[0050] In certain embodiments, single-chain Fv (scFv) antibodies are contemplated. For example, kappabodies (Ill et al., Prot. Eng. 10:949-57, 1997); minibodies (Martin et al., EMBO J 13:5305-9, 1994); diabodies (Holliger et al., PNAS 90:6444-8, 1993); or Janusins ​​(Traunecker et al., EMBO J 10:3655-59, 1991; and Traunecker et al., Int. J. Cancer Suppl. 7:51-52, 1992) can be prepared using standard molecular biology techniques following the teachings of the present application for selecting antibodies with the desired specificity.

[0051] Single-chain Fv (scFv) polypeptides are covalently linked VH::VL heterodimers expressed from gene fusions containing VH and VL-encoding genes connected by a peptide-encoding linker. Huston et al. (PNAS USA. 85(16):5879-5883, 1988). Numerous methods have been described for identifying chemical structures for converting naturally aggregated but chemically separated light and heavy polypeptide chains from antibody V regions into scFv molecules that fold into a three-dimensional structure substantially similar to that of an antigen-binding site. See, for example, U.S. Patent Nos. 5,091,513 and 5,132,405 to Huston et al. and U.S. Patent No. 4,946,778 to Ladner et al.

[0052] In certain embodiments, the antibodies or antigen-binding fragments described herein are in the form of "diabodies." Diabodies are multimers of polypeptides, each of which contains a first domain comprising an immunoglobulin light chain binding region and a second domain comprising an immunoglobulin heavy chain binding region, the two domains being linked (e.g., by a peptide linker) but unable to associate with each other to form an antigen-binding site; the antigen-binding site is formed by the association of a first domain of one polypeptide in the multimer with a second domain of another polypeptide in the multimer (WO 94 / 13804). A dAb fragment of an antibody consists of a VH domain (Ward et al., Nature 341:544-546, 1989). Diabodies and other multivalent or multispecific fragments can be constructed, for example, by gene fusion (see WO 94 / 13804; and Holliger et al., PNAS USA. 90:6444-6448, 1993).

[0053] Also included are minibodies comprising scFvs linked to CH3 domains (see Hu et al., Cancer Res. 56:3055-3061, 1996). See also Ward et al., Nature. 341:544-546, 1989; Bird et al., Science. 242:423-426, 1988; Huston et al., PNAS USA. 85:5879-5883, 1988); PCT / US92 / 09965; WO94 / 13804; and Reiter et al., Nature Biotech. 14:1239-1245, 1996.

[0054] Where bispecific antibodies are used, they may be conventional bispecific antibodies, which can be produced in a variety of ways (Holliger and Winter, Current Opinion Biotechnol. 4:446-449, 1993), for example, prepared chemically or from hybridomas, or may be any of the bispecific antibody fragments mentioned above.

[0055] Bispecific diabodies, in contrast to bispecific whole antibodies, can also be particularly useful because they can be easily constructed and expressed in E. coli. Diabodies (and many other polypeptides, such as antibody fragments) of suitable binding specificities can be easily selected using phage display (WO94 / 13804) from libraries. A library can be created in which one arm of the diabody is held constant, for example, with specificity directed against antigen X, and then the other arm is varied to select antibodies of suitable specificity. Bispecific whole antibodies can be generated by a number of methods, including knob-into-hole engineering (Brinkman & Kontermann, mAbs 9:182-212, 2017) (Ridgeway et al., Protein Eng. 9:616-621, 1996).

[0056] In certain embodiments, the antibodies or antigen-binding fragments described herein are in the form of a UniBody®. A UniBody® is an IgG4 antibody with the hinge region removed (see GenMab Utrecht, The Netherlands; see also US20090226421). This antibody technology creates a stable, smaller antibody format with an expected longer therapeutic window than current small antibody formats. IgG4 antibodies are considered inert and thus do not interact with the immune system. Fully human IgG4 antibodies can be engineered by removing the hinge region of the antibody to obtain half-molecule fragments with different stability characteristics compared to the corresponding intact IgG4 (GenMab, Utrecht). Halving the IgG4 molecule leaves only one region on the UniBody® that can bind to the cognate antigen (e.g., disease target); therefore, the UniBody® binds monovalently to only one site on the target cell. For certain cancer cell surface antigens, this monovalent binding will not stimulate cancer cells to grow as may be seen using a bivalent antibody with the same antigen specificity; therefore, UniBody® technology may offer a treatment option for some types of cancer that may be refractory to treatment with conventional antibodies. The small size of UniBody® may be of great benefit in treating some forms of cancer, but also allows for better distribution of the molecule to larger solid tumors, potentially increasing efficacy.

[0057] In certain embodiments, the antibodies and antigen-binding fragments described herein are in the form of Nanobodies. Nanobodies are encoded by a single gene and are efficiently produced in almost all prokaryotic and eukaryotic hosts, such as Escherichia coli (see U.S. Pat. No. 6,765,087), molds (e.g., Aspergillus or Trichoderma), and yeasts (e.g., Saccharomyces, Kluyveromyces, Hansenula, or Pichia (see U.S. Pat. No. 6,838,254). The production process is scalable, and multi-kilogram quantities of Nanobodies have been produced. Nanobodies can be formulated as a ready-to-use solution with a long shelf life. The Nanoclone method (see WO 06 / 079372) is a proprietary method for generating Nanobodies against desired targets based on automated high-throughput selection of B cells.

[0058] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of aptamers (see, e.g., Ellington et al., Nature. 346, 818-22, 1990; and Tuerk et al., Science. 249, 505-10, 1990, which are incorporated by reference). Examples of aptamers include nucleic acid aptamers (e.g., DNA aptamers, RNA aptamers) and peptide aptamers. Nucleic acid aptamers generally refer to nucleic acid species that are engineered through repeated rounds of in vitro selection or equivalent methods, such as SELEX (systematic evolution of ligands by exponential enrichment), to bind to various molecular targets, such as small molecules, proteins, nucleic acids, and even cells, tissues, and organisms. See, e.g., U.S. Patent Nos. 6,376,190 and 6,387,620, which are incorporated by reference.

[0059] Peptide aptamers typically contain a variable peptide loop attached at both ends to a protein scaffold, typically a dual structural constraint that increases the binding affinity of the peptide aptamer to levels comparable to those of antibodies (e.g., in the nanomolar range). In certain embodiments, the variable loop length may be comprised of approximately 10 to 20 amino acids (including all integers therebetween), and the scaffold may comprise any protein with good solubility and compaction properties. Certain exemplary embodiments utilize the bacterial protein thioredoxin-A as the scaffold protein, with the variable loop inserted into the reducing active site (the -Cys-Gly-Pro-Cys-loop in the wild-type protein), and two cysteine ​​side chains capable of forming disulfide bridges. Methods for identifying peptide aptamers are described, for example, in U.S. Patent Application Publication No. 2003 / 0108532, which is incorporated by reference. Peptide aptamer selection can be performed using different systems known in the art, including the yeast two-hybrid system.

[0060] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of avimers. Avimers refer to multimeric binding proteins or peptides engineered using in vitro exon shuffling and phage display. Multiple binding domains are linked, resulting in greater affinity and specificity compared to single-epitope immunoglobulin domains. See, for example, Silverman et al., Nature Biotechnology. 23:1556-1561, 2005; U.S. Patent No. 7,166,697; and U.S. Patent Application Nos. 2004 / 0175756, 2005 / 0048512, 2005 / 0053973, 2005 / 0089932, and 2005 / 0221384, which are incorporated by reference.

[0061] In some embodiments, the antibody or antigen-binding fragment described herein is in the form of an Adnectin. Adnectins refer to a class of targeted biologics derived from human fibronectin, an abundant extracellular protein that naturally binds to other proteins. See, for example, U.S. Patent Application Nos. 2007 / 0082365; 2008 / 0139791; and 2008 / 0220049, which are incorporated by reference. Adnectins typically consist of a natural fibronectin backbone and multiple targeting domains of specific parts of human fibronectin. The targeting domains can be engineered to enable Adnectins to specifically recognize IL-18BP polypeptides or epitopes thereof.

[0062] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of anticalins. Anticalins refer to a class of antibody mimics typically synthesized from human lipocalins, a family of binding proteins with hypervariable loop regions supported by a structurally rigid framework. See, for example, U.S. Patent Application No. 2006 / 0058510. Anticalins typically have a size of approximately 20 kDa. Anticalins can be characterized by a barrel structure formed by eight antiparallel β-strands (a stable β-barrel scaffold) connected in pairs by four peptide loops and attached α-helices. In certain aspects, conformational deviations are created in the hypervariable loop regions to achieve specific binding. See, for example, Skerra, FEBS J. 275:2677-83, 2008, incorporated by reference.

[0063] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of designed ankyrin repeat proteins (DARPins). DARPins comprise a class of non-immunoglobulin proteins that may offer advantages over antibodies in target binding in drug discovery and development. Among other applications, DARPins are ideally suited for in vivo imaging or delivery of toxins or other therapeutic payloads due to their favorable molecular properties, including small size and high stability. Low-cost production in bacteria and rapid generation of many target-specific DARPins make the DARPin approach useful for drug discovery. In addition, DARPins can be easily produced in multispecific formats, offering the potential to target effector DARPins to specific organs or target multiple receptors with a single molecule composed of several DARPins. See, e.g., Stumpp et al., Curr Opin Drug Discov Devel. 10:153-159, 2007; U.S. Patent Application No. 2009 / 0082274; and PCT / EP2001 / 10454, which are incorporated by reference.

[0064] Also included are heavy chain dimers, such as antibodies from camelids and sharks. Camelid and shark antibodies contain two homodimeric pairs of V-like and C-like domains (neither of which has a light chain). The V of heavy chain dimers in camelids is H Because the heavy chains do not need to make hydrophobic interactions with the light chains, the regions of the heavy chains that normally contact the light chains are changed to hydrophilic amino acid residues in camelids. The VH domains of heavy chain dimer IgGs are called VHH domains. Shark Ig-NARs contain a homodimer of one variable domain (called the V-NAR domain) and five C-like constant domains (C-NAR domains).

[0065] In camelids, the diversity of the antibody repertoire is determined by complementarity-determining regions (CDRs) 1, 2, and 3 in the VH or VHH regions. CDR3 in camelid VHH regions is characterized by its relatively long length, averaging 16 amino acids (Muyldermans et al., 1994, Protein Engineering 7(9):1129). This contrasts with the CDR3 regions of antibodies from many other species. For example, the CDR3 of mouse VH has an average of 9 amino acids. Libraries of camelid-derived antibody variable regions maintain the in vivo diversity of camelid variable regions and can be generated, for example, by the method disclosed in U.S. Patent Application No. 20050037421, published February 17, 2005.

[0066] In certain embodiments, the antibody or antigen-binding fragment thereof is humanized. These embodiments generally refer to chimeric molecules, prepared using recombinant techniques, having an antigen-binding site derived from an immunoglobulin from a non-human species and the remaining immunoglobulin structure of the molecule based on the structure and / or sequence of a human immunoglobulin. The antigen-binding site may comprise either a complete variable domain fused onto a constant domain, or only CDRs (whole or part) grafted onto appropriate framework regions in the variable domain. The epitope-binding site may be wild-type or modified by one or more amino acid substitutions. This eliminates the constant region as an immunogen in human individuals, although the possibility of an immune response to the foreign variable region remains (LoBuglio et al., PNAS USA 86:4220-4224, 1989; Queen et al., PNAS USA. 86:10029-10033, 1988; Riechmann et al., Nature. 332:323-327, 1988). Exemplary methods for antibody humanization include those described in US Pat. No. 7,462,697.

[0067] Another approach focuses not only on providing human-derived constant regions, but also on modifying the variable regions and reshaping them as closely as possible to human form. Both heavy and light chain variable regions are known to contain three complementarity-determining regions (CDRs) that vary in response to the epitope in question and determine binding ability, flanked by four framework regions (FRs) that are relatively conserved in a given species and presumably provide a scaffold for the CDRs. When a non-human antibody is prepared for a specific epitope, the variable region can be reshaped or humanized by grafting CDRs derived from the non-human antibody onto the FRs present in the modified human antibody. Application of this approach to various antibodies has been reviewed by Sato et al., Cancer Res. 53:851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J. Immunol. 148:1149-1154, 1992. In some embodiments, a humanized antibody retains all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs from the mouse antibody). In some embodiments, only a portion of the CDR sequences are transplanted from a non-human antibody (Bowers et al., J. Biol. Chem. 288:7688-7696, 2013).In certain embodiments, a humanized antibody has one or more CDRs (one, two, three, four, five, six) that are altered relative to the original antibody, also referred to as one or more CDRs "derived from" one or more CDRs of the original antibody.

[0068] In certain embodiments, the antibody is a chimeric antibody. In this regard, a chimeric antibody is composed of an antigen-binding fragment of an antibody operably linked or otherwise fused to a heterologous Fc portion of a different antibody. In certain embodiments, the Fc domain or heterologous Fc domain is of human origin. In certain embodiments, the Fc domain or heterologous Fc domain is of murine origin. In other embodiments, the heterologous Fc domain can be from an Ig class different from that of the parent antibody, including IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), and IgM. In further embodiments, the heterologous Fc domain can be composed of CH2 and CH3 domains derived from one or more of the different Ig classes. As described above for humanized antibodies, an antigen-binding fragment of a chimeric antibody can include only one or more of the CDRs of an antibody described herein (e.g., one, two, three, four, five, or six CDRs of an antibody described herein), or can include the entire variable domain (VL, VH, or both).

[0069] The term "bond" refers to a direct association between two molecules, for example, due to covalent, electrostatic, hydrophobic, and ionic and / or hydrogen-bonding interactions, including, for example, salt bridges and water bridges.

[0070] By "coding sequence" is meant any nucleic acid sequence that contributes to the code for the polypeptide product of a gene. In contrast, the term "non-coding sequence" refers to any nucleic acid sequence that does not directly contribute to the code for the polypeptide product of a gene.

[0071] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer, or group of elements or integers, but not the exclusion of any other element or integer or group of elements or integers.

[0072] "Consisting of" means including, but not limited to, everything that follows the phrase "consisting of." Thus, the phrase "consisting of" indicates that the listed elements are required or essential, and that no other elements will be present. By "consisting essentially of," it is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or function specified in this disclosure for the listed elements. Thus, the phrase "consisting essentially of" indicates that the listed elements are required or essential, but that other elements are optional and may or may not be present depending on whether they substantially affect the activity or function of the listed elements.

[0073] The term "effector function" in the context of an antibody, or "ADCC effector function," refers to the antibody's ability to interact with other arms of the immune system, including, for example, activation of the classical complement pathway or through Fc receptor engagement. The complement-dependent pathway is primarily driven by the interaction of C1q with the C1 complex and the clustered antibody Fc domain. Antibody-dependent cellular cytotoxicity (ADCC) is primarily driven by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG, which itself binds to the target cell. Fc receptors (FcRs) are key immunoregulatory receptors that connect antibody-mediated (humoral) immune responses to cellular effector functions. Receptors for all classes of immunoglobulins have been identified, including FcγR (IgG), FcεRI (IgE), FcαRI (IgA), FcμR (IgM), and FcδR (IgD). There are at least three receptor classes for human IgG found on leukocytes: CD64 (FcγRI), CD32 (FcγRIIa, FcγRIIb, and FcγRIIc), and CD16 (FcγRIIIa and FcγRIIIb). FcγRI is a high-affinity receptor (nanomolar range K D ), whereas FcγRII and FcγRIII have low to intermediate affinity (micromolar range K D Upon Fc binding, signaling pathways are triggered, leading to the secretion of various substances, such as lytic enzymes, perforin, granzymes, and tumor necrosis factor, which mediate the destruction of target cells. The level of ADCC effector function varies for human IgG subtypes. This is dependent on the allotype and the specific FcvR, but in simple terms, ADCC effector function is "high" for human IgG1 and IgG3, and "low" for IgG2 and IgG4.

[0074] The terms "endotoxin-free" or "substantially endotoxin-free" generally refer to compositions, solvents, and / or containers containing only trace amounts of endotoxin (e.g., amounts that have no clinically adverse physiological effects in a subject), and preferably undetectable amounts of endotoxin. Endotoxins are toxins associated with certain microorganisms, such as bacteria, typically gram-negative bacteria, although endotoxins can also be found in gram-positive bacteria, such as Listeria monocytogenes. The most common endotoxins are lipopolysaccharides (LPS) or lipooligosaccharides (LOS), found in the outer membrane of various gram-negative bacteria, which represent central pathogenic features in the ability of these bacteria to cause disease. Small amounts of endotoxin in humans can produce fever, a drop in blood pressure, and activation of inflammation and coagulation, among other adverse physiological effects.

[0075] Therefore, in pharmaceutical production, it is often desirable to remove most or all traces of endotoxin from the drug product and / or drug container, since even small amounts can cause adverse effects in humans. Dehydrogenation ovens can be used for this purpose, since temperatures above 300°C are typically required to decompose most endotoxins. For example, based on primary packaging materials such as syringes or vials, a glass temperature of 250°C in combination with a 30-minute hold time is often sufficient to achieve a 3-log reduction in endotoxin levels. Other methods of removing endotoxin are contemplated, including, for example, chromatography and filtration methods described herein and known in the art.

[0076] Endotoxin can be detected using routine techniques known in the art. For example, the Limulus amebocyte lysis assay utilizes blood from horseshoe crabs and is a highly sensitive assay for detecting the presence of endotoxin. In this test, very low levels of LPS can cause detectable clotting of the Limulus lysate due to a powerful enzyme cascade that amplifies this reaction. Endotoxin can also be quantified by enzyme-linked immunosorbent assay (ELISA). To be substantially endotoxin-free, endotoxin levels can be less than about 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.08, 0.09, 0.1, 0.5, 1.0, 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, or 10 EU / mg of active compound. Typically, 1 ng of lipopolysaccharide (LPS) corresponds to approximately 1-10 EU.

[0077] The term "epitope" includes any determinant, preferably a polypeptide determinant, capable of specific binding to an immunoglobulin or T-cell receptor. An epitope comprises the region of an antigen bound by an antibody. In certain embodiments, epitopic determinants include chemically active surface groupings of molecules, such as amino acids, sugar side chains, phosphoryls, or sulfonyls, and in certain embodiments, may have specific three-dimensional structural characteristics and / or specific charge characteristics. Epitopes can be continuous or discontinuous with respect to the primary structure of an antigen, e.g., an IL-18BP polypeptide. In certain embodiments, an epitope comprises, consists of, or consists essentially of about, at least about, or no more than about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous amino acids (i.e., a linear epitope) or non-contiguous amino acids (i.e., a conformational epitope) of a reference sequence (e.g., see Table B1) or target molecule described herein.

[0078] An "epitope" includes that portion of an antigen or other macromolecule that interacts with the variable region binding pocket of a binding protein and is capable of forming a binding interaction. Such a binding interaction may manifest as intermolecular contact with one or more amino acid residues of a CDR. Antigen binding may involve a CDR3 or CDR3 pair. An epitope can be a linear peptide sequence (i.e., "continuous") or can be composed of a discontinuous amino acid sequence (i.e., "conformational" or "discontinuous"). A binding protein can recognize more than one amino acid sequence; therefore, an epitope can define more than one distinct amino acid sequence. Epitopes recognized by binding proteins can be determined by peptide mapping and sequence analysis techniques well known to those skilled in the art. A "cryptic epitope" or "cryptic binding site" is an epitope or binding site of a protein sequence that is not exposed or is substantially protected from recognition in an unmodified polypeptide, but that can be recognized by a binding protein in a denatured or proteolytic polypeptide. The amino acid sequence that is not exposed or only partially exposed in the unmodified polypeptide structure is a potential cryptic epitope.If an epitope is not exposed or only partially exposed, then it is likely to be buried inside the polypeptide.Candidate cryptic epitopes can be identified, for example, by examining the three-dimensional structure of unmodified polypeptide.

[0079] The term "half maximum effective concentration" or "EC 50 " refers to the concentration of an agent (e.g., antibody) described herein that induces a response halfway between baseline and maximum after some specified exposure time; EC of a graded dose-response curve. 50 Therefore, EC50 represents the concentration of a compound at which 50% of its maximal effect is observed. EC50 also represents the plasma concentration required to obtain 50% of the maximal effect in vivo. Similarly, "EC 90 " refers to the concentration of an agent or composition at which 90% of its maximum effect is observed. 90 " is "EC 50and the Hill slope, or it can be determined directly from the data using routine knowledge in the art. In some embodiments, the EC 50 is less than about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, or 500 nM. In some embodiments, the agent has an EC50 value of about 1 nM or less.

[0080] "Immune response" refers to any immunological response derived from the immune system, including responses from the cellular and humeral, innate, and adaptive immune systems. Exemplary cellular immune cells include, for example, lymphocytes, macrophages, T cells, B cells, NK cells, neutrophils, eosinophils, dendritic cells, mast cells, monocytes, and all subsets thereof. Cellular responses include, for example, effector functions, cytokine release, phagocytosis, efferocytosis, translocation, trafficking, proliferation, differentiation, activation, suppression, cell-cell interactions, apoptosis, and the like. Humoral responses include, for example, IgG, IgM, IgA, IgE, responses, and their corresponding effector functions.

[0081] The "half-life" of a drug, such as an antibody, can refer to the time it takes for the drug to lose half of its pharmacological, physiological, or other activity, relative to such activity at the time of administration to the serum or tissues of an organism, or relative to any other defined time point. "Half-life" can also refer to the time it takes for the amount or concentration of the drug to decrease by half of the starting amount administered to the serum or tissues of an organism, relative to such amount or concentration at the time of administration to the serum or tissues of an organism, or relative to any other defined time point. Half-life can be measured in serum and / or in any one or more selected tissues.

[0082] The terms "modulating" and "altering" include "increasing," "enhancing," or "stimulating," as well as "decreasing," "reducing," or "inhibiting," typically in a statistically significant or physiologically significant amount or degree compared to a control. An "increased," "stimulated," or "enhanced" amount is typically a "statistically significant" amount and can include an increase that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100-fold or more (e.g., 500, 1000-fold) (including all integers and ranges therebetween, e.g., 1.5, 1.6, 1.7, 1.8) than the amount produced by no composition (e.g., absence of agent) or a control composition. A "decreased" or "reduced" or "inhibited" amount is typically a "statistically significant" amount and can include a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% decrease (including all integers and ranges therebetween) in the amount produced by no composition (e.g., absence of agent) or a control composition. Examples of comparisons and "statistically significant" amounts are described herein.

[0083] The terms "polypeptide," "protein," and "peptide" are used interchangeably and refer to a polymer of amino acids, not limited to any particular length. The term "enzyme" includes polypeptide or protein catalysts. This term includes modifications, such as myristoylation, sulfation, glycosylation, phosphorylation, and the addition or deletion of a signal sequence. The term "polypeptide" or "protein" refers to one or more chains of amino acids, where each chain contains amino acids covalently linked by peptide bonds, and where the polypeptide or protein can contain multiple chains noncovalently and / or covalently linked together by peptide bonds, including native proteins, i.e., proteins produced by natural cells and specific non-recombinant cells, or genetically engineered or recombinant cells, and includes molecules having the amino acid sequence of a native protein or molecules with deletions from, additions to, and / or substitutions of one or more amino acids from the native sequence. In certain embodiments, a polypeptide is a "recombinant" polypeptide produced by a recombinant cell containing one or more recombinant DNA molecules, which are typically made from heterologous polynucleotide sequences or combinations of polynucleotide sequences not otherwise found in the cell.

[0084] The terms "polynucleotide" and "nucleic acid" include mRNA, RNA, cRNA, cDNA, and DNA. The terms refer to polymeric forms of nucleotides, typically at least 10 bases in length, either ribonucleotides or deoxynucleotides, or modified forms of either type of nucleotide. The terms include single- and double-stranded forms of DNA. The terms "isolated DNA," "isolated polynucleotide," and "isolated nucleic acid" refer to molecules isolated free of total genomic DNA of a particular species. Thus, an isolated DNA segment encoding a polypeptide refers to a DNA segment containing one or more coding sequences that is isolated substantially free of, or purified free of, total genomic DNA of the species from which the DNA segment is obtained. Also included are non-coding polynucleotides (e.g., primers, probes, oligonucleotides) that do not encode a polypeptide. Also included are recombinant vectors, including, for example, expression vectors, viral vectors, plasmids, cosmids, phagemids, phages, viruses, and the like.

[0085] Additional coding or non-coding sequences may, but need not, be present within the polynucleotides described herein, and polynucleotides may, but need not, be linked to other molecules and / or supporting materials. Thus, polynucleotides or expressible polynucleotides, regardless of the length of the coding sequence itself, may be combined with other sequences, for example, expression control sequences.

[0086] "Expression control sequences" include regulatory sequences of nucleic acids or corresponding amino acids, such as promoters, leaders, enhancers, introns, recognition motifs for RNA or DNA binding proteins, polyadenylation signals, terminators, internal ribosome entry sites (IRES), secretion signals, subcellular localization signals, and the like, which can affect the transcription or translation of coding sequences in host cells, or the intracellular or cellular location. Exemplary expression control sequences are described in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990).

[0087] A "promoter" is a DNA regulatory region capable of binding RNA polymerase in a cell and initiating transcription of a downstream (3' direction) coding sequence. As used herein, a promoter sequence is bounded at its 3' end by a transcription initiation site, extends upstream (5' direction), and includes the minimum number of bases or elements necessary to initiate transcription at levels detectable above background. A transcription initiation site (conveniently defined by mapping with nuclease S1) can be found within the promoter sequence, as well as protein binding domains (consensus sequences) responsible for RNA polymerase binding. Eukaryotic promoters often, but not always, contain "TATA" and "CAT" boxes. Prokaryotic promoters contain Shine-Dalgarno sequences in addition to the -10 and -35 consensus sequences.

[0088] Numerous promoters, including constitutive, inducible, and repressible promoters from a variety of different sources, are well known in the art. Representative sources include, for example, viral, mammalian, insect, plant, yeast, and bacterial cell types. Suitable promoters from these sources are readily available online or based on publicly available sequences, for example, from repositories such as the ATCC, as well as other commercial or individual sources, or can be synthetically produced. Promoters can be unidirectional (i.e., initiate transcription in one direction) or bidirectional (i.e., initiate transcription in either the 3' or 5' direction). Non-limiting examples of promoters include, for example, the T7 bacterial expression system, the pBAD (araA) bacterial expression system, the cytomegalovirus (CMV) promoter, the SV40 promoter, and the RSV promoter. Inducible promoters include the Tet system (U.S. Pat. Nos. 5,464,758 and 5,814,618), the ecdysone-inducible system (No et al., Proc. Natl. Acad. Sci. (1996) 93(8):3346-3351; the T-REx™ system (Invitrogen, Carlsbad, CA), LacSwitch® (Stratagene, San Diego, CA), and the Cre-ERT tamoxifen-inducible recombinase system (Indra et al. Nuc. Acid. Res. (1999) 27(22):4324-4327; Nuc. Acid. Res. (2000) 28(23):e99; U.S. Pat. No. 7,112,715; and Kramer & Fussenegger, Methods Mol. Biol. (2005) 308:123-144), or any promoter known in the art suitable for expression in the desired cells.

[0089] An "expressible polynucleotide" includes a cDNA, RNA, mRNA, or other polynucleotide that comprises at least one coding sequence and, optionally, at least one expression control sequence, e.g., a transcriptional and / or translational regulatory element, and that is capable of expressing an encoded polypeptide upon introduction into a cell, e.g., a cell in a subject.

[0090] Various viral vectors that can be used to deliver expressible polynucleotides include adenovirus vectors, herpesvirus vectors, vaccinia virus vectors, adeno-associated virus (AAV) vectors, and retrovirus vectors. In some cases, the retrovirus vector is a derivative of a murine or avian retrovirus, or a lentivirus vector. Examples of retrovirus vectors that can insert a single foreign gene include, but are not limited to, Moloney murine leukemia virus (MoMuLV), Harvey murine sarcoma virus (HaMuSV), mouse mammary tumor virus (MuMTV), SIV, BIV, HIV, and Rous sarcoma virus (RSV). Many additional retroviral vectors can incorporate multiple genes. All of these vectors can transfer or incorporate genes for selectable markers, allowing transduced cells to be identified and generated. For example, a polypeptide sequence of interest can be inserted into the viral vector along with another gene encoding a ligand for a receptor on a specific target cell, making the vector target-specific. Retroviral vectors can be made target-specific, for example, by inserting a polynucleotide encoding a protein. An exemplary targeting can be achieved by using an antibody to target retroviral vectors. Those skilled in the art will know or can easily identify, without undue experimentation, the specific polynucleotide sequence that can be inserted into retroviral genome to enable target-specific delivery of retroviral vectors.

[0091] In certain embodiments, the expressible polynucleotide is a modified RNA or mRNA polynucleotide, e.g., a non-natural RNA analog. In certain embodiments, the modified RNA or mRNA polypeptide contains one or more modified or non-natural bases, e.g., a nucleotide base other than adenine (A), guanine (G), cytosine (C), thymine (T), and / or uracil (U). In some embodiments, the modified mRNA contains one or more modified or non-natural internucleotide linkages. Expressible RNA polynucleotides for delivering encoded therapeutic polypeptides are described, for example, in Kormann et al., Nat Biotechnol. 29:154-7, 2011; and U.S. Patent Application Nos. 2015 / 0111248; 2014 / 0243399; 2014 / 0147454; and 2013 / 0245104, which are incorporated by reference in their entireties.

[0092] The term "isolated" polypeptide or protein, as referred to herein, means that the subject protein (1) is free from at least some other proteins with which it would typically be found in nature; (2) is essentially free from other proteins from the same source, e.g., from the same species; (3) is expressed by cells from a different species; (4) is separated from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials with which it is naturally associated; (5) is not associated (by covalent or non-covalent interactions) with portions of proteins with which it is naturally associated; (6) is operably associated (by covalent or non-covalent interactions) with polypeptides with which it is not naturally associated; or (7) does not occur in nature. Such isolated proteins may be encoded by genomic DNA, cDNA, mRNA, or other RNA, may be of synthetic origin, or any combination thereof. In certain embodiments, an isolated protein is substantially free from proteins or polypeptides found in its natural environment or other contaminants that would interfere with its use (therapeutic, diagnostic, preventative, research, or otherwise).

[0093] In certain embodiments, the purity of any given agent (e.g., an antibody) in a composition can be defined. For example, a particular composition may include an agent that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% pure on a protein basis or weight-by-weight basis, including, for example, all decimal points measured and ranges therebetween, and is not limited by any means by high performance liquid chromatography (HPLC), a well-known form of column chromatography frequently used in biochemistry and analytical chemistry to separate, identify, and quantify compounds.

[0094] The term "reference sequence" generally refers to a nucleic acid coding sequence, or amino acid sequence, to which another sequence is compared. All polypeptide and polynucleotide sequences described herein are included as reference sequences, including those described by name and in the tables and sequence listing.

[0095] Certain embodiments include biologically active "variants" and "fragments" of the polypeptides (e.g., antibodies) described herein, as well as polynucleotides encoding the same. "Variant" includes one or more substitutions, additions, deletions, and / or insertions relative to a reference polypeptide or polynucleotide (see, e.g., Tables and Sequence Listing). A variant polypeptide or polynucleotide, as described herein, comprises an amino acid or nucleotide sequence with at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity or similarity or homology to a reference sequence and substantially retains the activity of the reference sequence. Also included are sequences that consist of a reference sequence or that differ from a reference sequence by the addition, deletion, insertion, or substitution of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or more amino acids or nucleotides, and that substantially retain the activity of the reference sequence. In certain embodiments, the additions or deletions include C-terminal and / or N-terminal additions and / or deletions.

[0096] The term "sequence identity" or, as used herein, includes, for example, "at least 50% identical sequence", refers to the degree to which sequences are identical on a nucleotide-by-nucleotide basis or on an amino acid-by-amino acid basis over a comparison window.Thus, "sequence identity percentage" can be calculated by: comparing two optimally aligned sequences over a comparison window; determining the number of positions where identical nucleic acid bases (e.g., A, T, C, G, I) or identical amino acid residues (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys and Met) occur in both sequences, resulting in the number of identical positions; dividing the number of identical positions by the total number of positions in the comparison window (i.e., window size), and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal alignment of sequences for aligning a comparison window can be performed by computerized implementations of algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Drive, Madison, Wis., USA), or by inspection and best alignment (i.e., resulting in the highest percentage of homology over the comparison window) generated by any of a variety of selected methods. Reference can also be made to the BLAST family of programs, for example, as disclosed by Altschul et al., Nucl. Acids Res. 25:3389, 1997.

[0097] The term "solubility" refers to the ability of an agent (e.g., an antibody) provided herein to dissolve in a liquid solvent to form a homogeneous solution. Solubility is typically expressed as a concentration by mass of solute per unit volume of solvent (e.g., g of solute per kg of solvent, g per dL (100 mL), mg / mL, etc.), molarity, molar concentration, molar fraction, or other similar descriptions of concentration. The maximum equilibrium amount of solute that can be dissolved per volume of solvent is the solubility of the solute in that solvent under specific conditions, including temperature, pressure, pH, and solvent properties. In certain embodiments, solubility is measured at physiological pH or other pHs, such as pH 5.0, pH 6.0, pH 7.0, pH 7.4, pH 7.6, pH 7.8, or pH 8.0 (e.g., about pH 5-8). In certain embodiments, solubility is measured in water or a physiological buffer, such as PBS or NaCl (with or without NaPO). In certain embodiments, solubility is measured at a relatively low pH (e.g., pH 6.0) and a relatively high salt (e.g., 500 mM NaCl and 10 mM NaPO). In certain embodiments, solubility is measured in a biological fluid (solvent), such as blood or serum. In certain embodiments, the temperature can be about room temperature (e.g., about 20, 21, 22, 23, 24, 25°C) or about body temperature (37°C). In certain embodiments, the agent has a solubility of at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 70, 80, 90 or 100 mg / ml at room temperature or 37°C.

[0098] A "subject" or "subject in need thereof" or a "patient" or "patient in need thereof" includes mammalian subjects, such as human subjects.

[0099] "Substantially" or "essentially" means nearly entirely or completely, for example, 95%, 96%, 97%, 98%, 99%, or more of a given amount of some substance.

[0100] By " statistically significant ", it means that the result is unlikely to occur by chance. Statistical significance can be determined by any method known in the art. Commonly used significance measures include p-value, which is the frequency or probability that the observed event will occur if the null hypothesis is true. If the obtained p-value is smaller than the significance level, then the null hypothesis is rejected. In simple cases, the significance level is defined as a p-value of 0.05 or less.

[0101] "Therapeutic response" refers to an improvement in symptoms (whether sustained or not) following administration of one or more therapeutic agents.

[0102] As used herein, the term "therapeutically effective amount," "therapeutic dose," "prophylactically effective amount," or "diagnostically effective amount" is the amount of an agent (e.g., an anti-IL-18BP antibody, an immunotherapeutic agent) required to elicit a desired biological response following administration.

[0103] As used herein, "treatment" of a subject (e.g., a mammal, such as a human) or cell is any type of intervention used in an attempt to alter the natural course of a disease. Treatment includes, but is not limited to, the administration of a pharmaceutical composition and can be performed either prophylactically or following the initiation of a pathological event or contact with a pathogenic agent. Also included is prophylactic treatment, which can be directed at reducing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate complete eradication, cure, or prevention of the disease or condition, or its associated symptoms.

[0104] The term "wild-type" refers to a gene or gene product (eg, a polypeptide) that is most frequently observed in a population and is thus arbitrarily designed as the "normal" or "wild-type" form of the gene.

[0105] Each embodiment herein should be applied mutatis mutandis to all other embodiments unless expressly stated otherwise.

[0106] Anti-IL-18BP antibody Certain embodiments include antibodies, and antigen-binding fragments thereof, that bind to IL-18BP. In some embodiments, the antibodies or antigen-binding fragments thereof modulate (e.g., interfere with, antagonize, inhibit) the binding of IL-18BP to its ligand, interleukin-18 (IL-18). In certain embodiments, the antibodies or antigen-binding fragments thereof bind to the complementarity-determining region V of IL-18BP. H CDR1, V H CDR2, and V H The heavy chain variable region (V H ), and complementarity-determining region V L CDR1, V L CDR2, and V L The light chain variable region (V L ) characterized by or including them. H Array, V H CDR1 sequence, V H CDR2 sequence, V H CDR3 sequence, V L Array, V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences are provided in Tables A1 and A2 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6]

[0107] Thus, in certain embodiments, the antibody or antigen-binding fragment thereof comprises a V complementarity determining region selected from Table A1. H CDR1 sequence, V H CDR2 sequence, and V H V containing the CDR3 sequence H sequences, and variants thereof that bind to IL-18BP, and V of the complementarity determining region selected from Table A1 L CDR1 sequence, V L CDR2 sequence, and V L V containing the CDR3 sequence L In certain embodiments, the antibody comprises the sequence V H CDR1 sequence, V H CDR2 sequence, and V H V containing the CDR3 sequence H Sequence, and V L CDR1 sequence, V L CDR2 sequence, and V L V containing the CDR3 sequence L sequences, in which the CDR sequences are all derived from one named antibody of Table A1 (e.g., SA01a).

[0108] In certain embodiments, the CDR sequences are as follows: V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 4 to 6, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 28 to 30, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 31 to 33, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 34 to 36, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 37 to 39, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 40 to 42, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 46 to 48, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 52 to 54, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 58 to 60, respectively; V H CDR1 sequence, V HCDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 64 to 66, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 70 to 72, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 109 to 111, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 124 to 126, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and VL CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 130 to 132, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 136 to 138, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 139 to 141, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 142 to 144, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 148 to 150, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 154 to 156, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 160 to 162, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 172 to 174, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 190 to 192, respectively; V HCDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 193 to 195, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 199 to 201, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 205 to 207, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 208-210, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 211 to 213, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 214-216, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 217 to 219, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; V H CDR1 sequence, V H CDR2 sequence, and V HThe CDR3 sequences comprise SEQ ID NOs: 223 to 225, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 229 to 231, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 232 to 234, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 235 to 237, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 241 to 243, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 244 to 246, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 247 to 249, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 250 to 252, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 253 to 255, respectively, and V L CDR1 sequence, VL CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 256 to 258, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 265 to 267, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 268-270, respectively; or V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 271 to 273, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences comprise SEQ ID NOs: 274 to 276, respectively.

[0109] In certain embodiments, the antibody or antigen-binding fragment thereof (e.g., a variant of the SA01a antibody or antigen-binding fragment thereof) comprises CDR consensus sequences, e.g., the VHCDR1, VHCDR2, and VHCDR3 sequences are TFX1X2X3X4X5H, IX6X7X8X9X, and TFX1X2X3X4X5H, respectively. 10 X 11 X 12 X 13 X 14 X 15 AQKFQG, and X 16 X 17 X 18 X 19 X 20 X 21 X 22 DY, and the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each contain X 23 X 24 X 25 X 26 X 27 X 28 X 29 X 30 WX 31 A, X 32 X 33X 34 X 35 X 36 X 37 X 38 , and QX 39 X 40 X 41 SFPYX 42 (See Table E11 for definition of "X" residues).

[0110] Also included are minor variants of the aforementioned CDRs. Exemplary variants bind to IL-18BP and have minor variations in any one or more of the individual CDRs, e.g., the V and VV variants described herein. H CDR1, V H CDR2, V H CDR3, V L CDR1, V L CDR2, and / or V L Any one or more of the CDR3 sequences may have a total of 1, 2, or 3 changes. Exemplary "changes" include amino acid substitutions, additions, and deletions.

[0111] Exemplary V H and V L The sequences are provided in Table A2 below. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0112] Thus, in certain embodiments, the antibody, or antigen-binding fragment thereof, binds to IL-18BP and is selected from Table A2. H Array and corresponding V LIn certain embodiments, V H includes a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, e.g., V H has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions. L includes a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, e.g., V L has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions. H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and V H In certain embodiments, the V region is derived from the same named antibody (e.g., SA01a antibody). H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and is derived from the same one named antibody (e.g., SA01a antibody) as the VH region, where no changes are present in the underlined CDRs of Table A2. Thus, the antibody comprises a VH region at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to each sequence from one named antibody (e.g., SA01a) of Table A2. H Array and V L Alternatively, the antibody may comprise a sequence, in which case the antibody comprises the CDRs of one of the named antibodies (e.g., SA01a) listed in Table A1.

[0113] In some embodiments, the V of the antibody or antigen-binding fragment H and VL is as follows: V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 286; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 288; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 292; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and VL comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 294; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 296; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 298; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 352; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 353, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 354; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 355, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 356; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 357, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 358; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 359, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 360; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 361, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 362; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 368; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 369, and V L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:370.

[0114] Also, variants thereof that bind to IL-18BP, such as the aforementioned V H Sequence and / or V L These are variants having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in any one or more framework regions of the sequence. Exemplary "changes" include amino acid substitutions, additions, and deletions.

[0115] As described above, the antibodies or antigen-binding fragments thereof described herein bind to IL-18BP. In certain embodiments, the antibodies or antigen-binding fragments thereof bind to human IL-18BP, cynomolgus monkey IL-18BP, and / or mouse IL-18BP, or regions, fragments, or epitopes thereof.

[0116] Human interleukin-18 binding protein, or IL-18BP, is encoded by the IL18BP gene (Gene ID: 10068; and UniProt: O95998) and has at least three isoforms. In some embodiments, an antibody of the present disclosure binds to isoform A of IL-18BP. In some embodiments, an antibody of the present disclosure binds to isoform B of IL-18BP. In some embodiments, an antibody of the present disclosure binds to both isoform A and isoform C of IL-18BP. In some embodiments, an antibody of the present disclosure binds to all isoforms of IL-18BP. It is an inhibitor of the early Th1 cytokine response and the pro-inflammatory cytokine IL-18. For example, IL-18BP binds to IL-18 and inhibits the binding of IL-18 to its receptor, thereby inhibiting IL-18-induced IFN-gamma production, among other IL-18 signaling activities. The amino acid sequences of human, cynomolgus monkey, and mouse IL-18BP isoforms are presented below in Table B1 (see also Figure 2 for alignment). [Table 3]

[0117] Thus, in certain embodiments, the antibody or antigen-binding fragment thereof binds to the mature IL-18BP sequence of Table B1, e.g., in the region excluding the signal peptide (underlined). In certain embodiments, the antibody or antigen-binding fragment thereof binds to an epitope comprising the IL-18-binding interface of mature IL-18BP. In some embodiments, the antibody or antigen-binding fragment thereof binds to an epitope comprising I97 and V153 according to UniProt:O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO:1, or I67 and V123 as set forth in SEQ ID NO:2).

[0118] In certain embodiments, the antibody binds to a conformational epitope of the mature IL-18BP sequence of SEQ ID NO: 372 (mature human isoform A). In exemplary embodiments, an antibody of the disclosure binds to at least two residues selected from the group consisting of K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO: 372. In exemplary embodiments, an antibody of the disclosure binds to residues K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO: 372. In exemplary embodiments, the antibody comprises a VH of a sequence with at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 333, and a VL of a sequence with at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 334. In exemplary embodiments, the VHCDR1, VHCDR2, and VHCDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and the VLCDR1, VLCDR2, and VLCDR3 sequences comprise SEQ ID NOs: 172-174, respectively.

[0119] In certain embodiments, the antibody binds to an epitope comprising the IL-18-binding interface of mature IL-18BP. The residues on IL-18BP that interact with IL-18 have been identified as follows: R61, Y69, S75, H79, T116, S119, and R131. In exemplary embodiments, the antibody of the present disclosure binds to residues R61, Y69, and R131, which are also recognized by IL-18. In exemplary embodiments, the VH has a sequence with at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 333, and the VL has a sequence with at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 334. In an exemplary embodiment, the antibody comprises VHCDR1, VHCDR2, and VHCDR3 sequences comprising SEQ ID NOs: 169-171, respectively, and VLCDR1, VLCDR2, and VLCDR3 sequences comprising SEQ ID NOs: 172-174, respectively.

[0120] In certain embodiments, the antibody binds to a linear epitope of the mature IL-18BP sequence of SEQ ID NO: 372 (mature human isoform A). In certain embodiments, the antibody has orthologous specificity or orthologous cross-reactivity to IL-18BP. For example, in certain embodiments, the antibody binds to human IL-18BP and cynomolgus IL-18BP, but does not bind (specifically or substantially) to mouse IL-18BP. In some embodiments, the antibody binds to human IL-18BP, cynomolgus IL-18BP, and mouse IL-18BP.

[0121] In certain embodiments, the antibody or antigen-binding fragment thereof has orthologous specificity or orthologous cross-reactivity to IL-18BP. For example, in certain embodiments, the antibody or antigen-binding fragment thereof binds to human IL-18BP and cynomolgus IL-18BP but does not bind (specifically or substantially) to mouse IL-18BP. In some embodiments, the antibody or antigen-binding fragment thereof binds to human IL-18BP, cynomolgus IL-18BP, and mouse IL-18BP (e.g., SA01a, SA51d, SA45a, SA54a, SA55a, SA56a, SA57a, SA59a, SA60a, SA61a, SA62a, SA63a, SA65a, SA73a, SA77a, SA64a, SA66a).

[0122] In some embodiments, the antibody or antigen-binding fragment thereof is directed to human IL-18BP by measuring the binding affinity between IL-18 and IL-18BP (K DIn some examples, the antibody or antigen-binding fragment thereof binds to human IL-18BP with a binding affinity of about 1 pM to about 10 pM to about 600 pM, or 65 pM, or about, at least about, or less than about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM, or optionally about 1 pM to about 10 pM to about 600 pM, or 65 pM. Approximately 600pM, 1pM to approximately 500pM, 1pM to approximately 400pM, 1pM to approximately 300pM, approximately 1pM to approximately 200pM, approximately 1pM to approximately 100pM, approximately 1pM to approximately 50pM, approximately 1pM to approximately 40pM, approximately 1pM to approximately 30pM, approximately 1pM to approximately 2 0pM, about 1pM to about 10pM, about 1pM to about 5pM, about 5pM to about 600pM, about 5pM to about 500pM, about 5pM to about 400pM, about 5pM to about 300pM, about 5pM to about 200pM, about 5pM to about 100pM, about 5pM to about 5 0 pM, about 5 pM to about 40 pM, about 5 pM to about 30 pM, about 5 pM to about 20 pM, about 5 pM to about 10 pM, about 10 pM to about 600 pM, about 10 pM to about 500 pM, about 10 pM to about 400 pM, about 10 pM to about 300 pM, about 10 pM to about 200 pM, about 10 pM to about 100 pM, about 10 pM to about 50 pM, about 10 pM to about 40 pM, about 10 pM to about 30 pM, about 10 pM to about 20 pM, or about 20 pM to about 600 pM, about 20 pM to about 500 pM, about 20 pM It binds with a binding affinity of about 20 pM to about 400 pM, about 20 pM to about 300 pM, about 20 pM to about 200 pM, about 20 pM to about 100 pM, about 20 pM to about 50 pM, about 20 pM to about 40 pM, about 20 pM to about 30 pM, or about 30 pM to about 600 pM, about 30 pM to about 500 pM, about 30 pM to about 400 pM, about 30 pM to about 300 pM, about 30 pM to about 200 pM, about 30 pM to about 100 pM, about 30 pM to about 50 pM, or about 30 pM to about 40 pM. Dmay be determined by the biolayer interference (BLI) assay described herein. For example, binding kinetics measurements may be performed on a ForteBio (now Sartorius) Octet RED96e instrument by loading the mAb onto an anti-human constant domain (AHC) biosensor (ForteBio) in 10x kinetics buffer consisting of PBS containing 0.1% BSA and 0.02% Tween 20 for 90-120 seconds, achieving spectral shift values ​​of 0.8-1.2 nm. Binding may then be performed in the presence of a two-fold dilution series of hIL-18BP and allowed to proceed for 90-120 seconds. Dissociation may be measured over 300-1200 seconds. The dilution series may start at 100 nM for weaker variants or 10 nM for the most potent mAb.

[0123] In some embodiments, the antibody, or antigen-binding fragment thereof, is an IL-18BP antagonist. In some examples, the antibody, or antigen-binding fragment thereof, antagonizes the binding and / or signaling activity between IL-18BP and its ligand, IL-18. In some embodiments, the antibody, or antigen-binding fragment thereof, antagonizes or reduces the binding and / or signaling activity between IL-18BP and IL-18 by about or at least about 10-1000% (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000% or more), e.g., in a cell-based assay. In some examples, an antagonistic anti-IL-18BP antibody or antigen-binding fragment thereof blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signal transduction, e.g., IL-18-mediated induction of IFN-γ, CXCL10, and TNFα. These functional activities may be measured by the assays disclosed herein. For example, an antibody may be incubated with IL-18BP (e.g., human IL-18BP) followed by the addition of IL-18 (e.g., recombinant human IL-18). The resulting solution may then be added to IL-18 reporter HEK293 cells. The cells respond to exogenously added IL-18 by expressing the NF-κB / AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene. The effect of the antibody may then be analyzed by its effect on the reporter cells compared with an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section of this specification. Another potential assay involves measuring the inhibition of IFNγ expression by anti-IL-18BP mAb in KG-1 cells. Briefly, IL-18BP may be blocked first using serial dilutions of the antibody, then IL-18 may be added to the mixture, and the mixture may be added to KG-1 cells and incubated.Secreted IFN-γ can then be measured according to standard means, such as ELISA, and the effect of the test antibody can be compared to that of an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section herein. Yet another possible assay involves incubating PBMCs with the test antibody, IL-12, and IL-18, and measuring IFN-γ and / or CCL2 by standard methods. The effect of the test antibody may be compared to that of an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section herein. A further assay involves incubation of NK cells with precomplexed hIL-18 / hIL-18BP, followed by the addition of IL-12, followed by the addition of serial dilutions of the test antibody. The effect of the test antibody is compared to that of an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section herein.

[0124] Certain embodiments include a method of screening an anti-IL-18BP antibody or antigen-binding fragment thereof for the ability to block or inhibit binding between IL-18 and IL-18BP, the method comprising: (a) determining the binding affinity of the antibody or antigen-binding fragment thereof to (i) IL-18BP alone, and (ii) an IL-18-reduced fusion protein, the IL-18-reduced fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween). (b) comparing the binding affinity of (i) with the binding affinity of (ii); and (c) identifying or selecting the antibody or antigen-binding fragment thereof as having the ability to block or inhibit binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii). Certain embodiments include (c) identifying or selecting the antibody or antigen-binding fragment thereof as having the ability to block or inhibit binding between IL-18 and IL-18BP if the binding affinity of (i) is about or at least about 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 200-fold, 300-fold, 400-fold, 500-fold, 600-fold, 700-fold, 800-fold, 900-fold, or 1000-fold or more stronger than the binding affinity of (ii). In certain embodiments, the IL-18 and IL-18BP are murine IL-18 and IL-18BP. In some embodiments, the IL-18 and IL-18BP are human IL-18 and IL-18BP. In some embodiments, the IL-18-reduced fusion protein comprises, from N- to C-terminus, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP.In certain embodiments, the low IL-18 fusion protein comprises an amino acid sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

[0125] For illustrative purposes only, binding interactions (e.g., binding affinity) between any combination of IL-18BP, IL-18 (e.g., low IL-18), and / or anti-IL-18BP antibodies, or antigen-binding fragments thereof, described herein, or binding / signaling between IL-18BP and IL-18, can be detected and quantified using a variety of routinely used methods, including Biacore® assays (e.g., coupled to a sensor chip using an appropriately tagged soluble reagent), FACS analysis using cells (either natural or recombinant) expressing IL-18BPr on their cell surface, immunoassays, fluorescent staining assays, ELISA assays, and microcalorimetry, e.g., ITC (isothermal titration calorimetry). Similarly, the functional properties of anti-IL-18BP antibodies can be evaluated using a variety of methods known to those skilled in the art, including affinity / binding assays (e.g., surface plasmon resonance, competitive inhibition assays); cytotoxicity assays, cell survival assays, cell proliferation or differentiation assays, and cancer cell and / or tumor growth inhibition using in vitro or in vivo models. Other assays may be used to test the ability of the antibodies described herein to modulate (e.g., inhibit) IL-18BP and / or IL-18-mediated responses. The antibodies described herein may be tested for in vitro and in vivo efficacy. Such assays may be performed using well-established protocols known to those skilled in the art (see, e.g., Current Protocols in Molecular Biology (Greene Publ. Assoc. Inc. & John Wiley & Sons, Inc., NY, NY); Current Protocols in Immunology (Edited by: John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strober 2001 John Wiley & Sons, NY, NY); or commercially available kits.

[0126] In certain embodiments, the Fc region of an antibody, or antigen-binding fragment thereof, comprises, consists of, or consists essentially of an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or a hybrid and / or variant thereof. In certain embodiments, the Fc region comprises, consists of, or consists essentially of an Fc from human IgG1 or IgG4 (see, e.g., Allberse and Schuurman, Immunology. 105:9-19, 2002), or a fragment or variant thereof.

[0127] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a variant or otherwise modified Fc region, including one with altered properties or biological activity compared to a wild-type Fc region. Examples of modified Fc regions include regions having a mutated sequence, e.g., by substitution, insertion, deletion, or truncation of one or more amino acids relative to the wild-type sequence, hybrid Fc polypeptides composed of domains from different immunoglobulin classes / subclasses, Fc polypeptides with altered glycosylation / sialylation patterns, and Fc polypeptides that are modified or derivatized, e.g., by biotinylation (see, e.g., U.S. Patent Application Publication No. 2010 / 0209424), phosphorylation, sulfation, etc., or any combination of the foregoing. Such modifications can affect the binding properties of the Fc region to one or more specific FcRs (e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, FcγRIIIb, FcRn), its pharmacokinetic properties (e.g., stability or half-life, bioavailability, tissue distribution, volume of distribution, concentration, elimination rate constant, elimination rate, area under the curve (AUC), clearance, C max , T max , C min, wander mutations), its immunogenicity, its complement fixation or activation, and / or CDC / ADCC / ADCP-related activity of the Fc region compared to the corresponding wild-type Fc sequence of an antibody or antigen-binding fragment thereof. Included are modified Fc regions of human and / or murine origin.

[0128] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a hybrid Fc region, e.g., an Fc region comprising a combination of Fc domains (e.g., hinge, CH2, CH3, CH4) from immunoglobulins of different species (e.g., human, mouse), different Ig classes, and / or different Ig subclasses. Also included are antibodies or antigen-binding fragments thereof comprising derivatized or otherwise modified Fc regions. In certain aspects, the Fc region is modified, e.g., by phosphorylation, sulfation, acylation, glycosylation, methylation, farnesylation, acetylation, amidation, and the like, relative to a wild-type or native Fc region. In certain embodiments, the Fc region comprises a wild-type or native glycosylation pattern, or alternatively, it comprises increased glycosylation relative to the native form, decreased glycosylation relative to the native form, or it is completely deglycosylated. As an example of an altered Fc glycoform, reduced glycosylation of the Fc region reduces binding to the C1q region of the first complement component C1, decreases ADCC-related activity, and / or decreases CDC-related activity. Certain embodiments thus employ deglycosylated or aglycosylated Fc regions. For exemplary production of aglycosylated Fc regions, see, e.g., WO 2005 / 047337. Another example of an Fc region glycoform is generated by substituting a cysteine ​​residue at position Q295 according to the Kabat et al. numbering system (see, e.g., U.S. Patent Application No. 2010 / 0080794). Certain embodiments include Fc regions in which approximately 80-100% of the glycoproteins in the Fc region comprise a mature core carbohydrate structure lacking fucose (see, e.g., U.S. Patent Application No. 2010 / 0255013). Some embodiments include Fc regions that are optimized by substitutions or deletions to reduce the level of fucosylation, e.g., to increase affinity for FcγRI, FcγRIa, or FcγRIIIa, and / or to improve phagocytosis by FcγRIIa-expressing cells (see U.S. Patent Applications Nos. 2010 / 0249382 and 2007 / 0148170).

[0129] As another example of a modified Fc glycoform, the Fc region of an antibody or antigen-binding fragment thereof may comprise oligomannose-type N-glycans, optionally having one or more of the following: increased ADCC effector activity, increased binding affinity for FcγRIIIA (and certain other FcRs), similar or increased binding specificity for a target of an IL-18BP polypeptide, similar or higher binding affinity for a target of an IL-18BP polypeptide, and / or similar or lower binding affinity for a mannose receptor, relative to a corresponding Fc region comprising complex-type N-glycans (see, e.g., U.S. Patent Application Publication No. 2007 / 0092521 and U.S. Patent No. 7,700,321). As another example, increased affinity of the Fc region for FcγRs has been achieved using engineered glycoforms generated by expression of the antibody in engineered or variant cell lines (see, e.g., Umana et al., Nat Biotechnol. 17:176-180, 1999; Davies et al., Biotechnol Bioeng. 74:288-294, 2001; Shields et al., J Biol Chem. 277:26733-26740, 2002; Shinkawa et al., 2003, J Biol Chem. 278:3466-3473, 2003; and U.S. Patent Application No. 2007 / 0111281). Certain Fc region glycoforms contain an increased proportion of N-glycosidically linked complex glycans that do not have the 1-position of fucose attached to the 6-position of the N-acetylglucosamine at the reducing end of the glycan (see, e.g., U.S. Patent Application Publication No. 2010 / 0092997). Certain embodiments may include an IgG Fc region that is glycosylated with at least one galactose moiety linked to each terminal sialic acid moiety by an α-2,6 linkage, optionally wherein the Fc region has greater anti-inflammatory activity than the corresponding wild-type Fc region (see, e.g., U.S. Patent Application Publication No. 2008 / 0206246).Some of these and related altered glycosylation approaches, as described herein, have produced substantial enhancements in the ability of the Fc region to selectively bind to FcRs, such as FcγRIII, mediate ADCC, and modify other properties of the Fc region.

[0130] Particular variant, fragment, hybrid, or otherwise modified Fc regions of antibodies or antigen-binding fragments thereof may have altered binding to one or more FcRs and / or corresponding changes in effector function compared to the corresponding wild-type Fc sequence (e.g., same species, same Ig class, same Ig subclass). For example, such Fc regions may have increased binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In other embodiments, variant, fragment, hybrid, or modified Fc regions may have decreased binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. Particular FcRs are described elsewhere herein.

[0131] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors relative to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations to increase binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors relative to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase effector function. In some embodiments, the antibody comprises an Fc domain selected from human IgG1 and human IgG3 comprising one or more mutations to increase effector function.

[0132] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with enhanced effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG1 and human IgG3, with one or more mutations to increase effector function. In some embodiments, the antibody is a partial blocking antibody comprising an Fc domain with enhanced effector activity. In some embodiments, the partial blocking antibody comprises an Fc domain selected from human IgG1 and human IgG3, with one or more mutations to increase effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with enhanced effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG1 or human IgG3, with one or more mutations to increase effector function.

[0133] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations to reduce binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce effector function. In some embodiments, the antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function.

[0134] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a partial blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the partial blocking antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function.

[0135] Specific examples of Fc variants with altered (e.g., increased, decreased) effector function / FcR binding are described, for example, in U.S. Patent Nos. 5,624,821 and 7,425,619; U.S. Patent Application Nos. 2009 / 0017023, 2009 / 0010921, and 2010 / 0203046; and WO2000 / 42072 and WO2004 / 016750. Specific examples include human Fc regions with one or more substitutions at positions 298, 333, and / or 334, e.g., S298A, E333A, and / or K334A (based on the EU index numbering of Kabat et al.), which have been shown to increase binding to the activating receptor FcγRIIIa and decrease binding to the inhibitory receptor FcγRIIb. These mutations can be combined to produce double and triple mutant variants with further improvements in FcR binding. Particular embodiments include the S298A / E333A / K334A triple mutant, which increases binding to FcγRIIIa, decreases binding to FcγRIIb, and increases ADCC (see, e.g., Shields et al., J. Biol. Chem. 276:6591-6604, 2001; and Presta et al., Biochem. Soc. Trans. 30:487-490, 2002). See also Umana et al. (supra); and engineered Fc glycoforms with increased FcR binding, as disclosed in U.S. Pat. No. 7,662,925. Some embodiments include an Fc region comprising one or more substitutions based on the EU index of Kabat et al. selected from 434S, 252Y / 428L, 252Y / 434S, and 428L / 434S (see U.S. Patent Application Nos. 2009 / 0163699 and 20060173170). Some embodiments include an Fc region comprising one or more substitutions based on the EU index of Kabat et al. selected from L234A, L235A, and G237A (see U.S. Patent Application No. 17 / 779,425). Some embodiments include an Fc region comprising substitutions at L234A and L235A based on the EU index of Kabat et al.Some embodiments include an Fc region comprising substitutions at L234A and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising substitutions at L235A and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising substitutions at L234A, L235A, and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising substitutions at L234A, L235A, and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising one or more substitutions selected from M252Y, S254T, and T256E based on the EU index of Kabat et al. Some embodiments include an Fc region comprising one or more substitutions selected from M428L and N434S based on the EU index of Kabat et al. In some embodiments, the Fc substitutions are made into an Fc domain selected from human IgG1, IgG2, IgG3, and IgG4. In some embodiments, the Fc substitutions are made into a human IgG1 Fc domain. In some embodiments, the Fc substitutions are made into a human IgG2 Fc domain. In some embodiments, the above-mentioned Fc substitutions are substitutions into a human IgG3 Fc domain. In some embodiments, the above-mentioned Fc substitutions are substitutions into a human IgG4 Fc domain. In some embodiments, antibodies of the present disclosure comprise an Fc substitution disclosed herein and a V that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to each sequence from one of the named antibodies of Table A2 (e.g., SA01a). H Array and V L The antibody comprises a sequence, in which case the antibody comprises the CDRs of one of the named antibodies (e.g., SA01a) listed in Table A1.

[0136] Certain variant, fragment, hybrid, or modified Fc regions may have altered effector function compared to the corresponding wild-type Fc sequence. For example, such Fc regions may have increased complement fixation or activation, increased Clq binding affinity, increased CDC-related activity, increased ADCC-related activity, and / or increased ADCP-related activity compared to the corresponding wild-type Fc sequence. In other embodiments, such Fc regions may have decreased complement fixation or activation, decreased Clq binding affinity, decreased CDC-related activity, decreased ADCC-related activity, and / or decreased ADCP-related activity compared to the corresponding wild-type Fc sequence. As merely one illustrative example, the Fc region may include deletions or substitutions in complement binding sites, such as C1q binding sites, and / or deletions or substitutions in ADCC sites. Examples of such deletions / substitutions are described, for example, in U.S. Patent No. 7,030,226. Many Fc effector functions, such as ADCC, can be assayed according to routine techniques in the art (see, e.g., Zuckerman et al., CRC Crit Rev Microbiol. 7:1-26, 1978). Useful effector cells for such assays include, but are not limited to, natural killer (NK) cells, macrophages, and other peripheral blood mononuclear cells (PBMCs). Alternatively, or additionally, certain Fc effector functions can be assessed in vivo by using animal models, such as those described in Clynes et al. PNAS. 95:652-656, 1998.

[0137] Certain variant hybrids or modified Fc regions may have altered stability or half-life compared to the corresponding wild-type Fc sequence. In certain embodiments, such Fc regions may have an increased half-life compared to the corresponding wild-type Fc sequence. In other embodiments, variant hybrids or modified Fc regions may have a decreased half-life compared to the corresponding wild-type Fc sequence. Half-life can be measured in vitro (e.g., under physiological conditions) or in vivo according to routine techniques in the art, such as radiolabeling, ELISA, or other methods. In vivo measurements of stability or half-life can be measured in one or more body fluids, including blood, serum, plasma, urine, or cerebrospinal fluid, or in a given tissue, such as liver, kidney, muscle, central nervous system tissue, bone, etc. As an example, modifications to an Fc region that alter its ability to bind to FcRn can alter its half-life in vivo. Non-limiting examples of assays for measuring in vivo pharmacokinetic properties (e.g., in vivo mean elimination half-life) and Fc modifications that alter their binding to FcRn are described, for example, in U.S. Pat. Nos. 7,217,797 and 7,732,570; and U.S. Patent Application Nos. US2010 / 0143254 and 2010 / 0143254.

[0138] Additional non-limiting examples of stability- or half-life-altering modifications include substitutions / deletions at one or more amino acid residues selected from 251-256, 285-290, and 308-314 of the CH2 domain, and 385-389 and 428-436 of the CH3 domain, according to the numbering system of Kabat et al. See U.S. Patent Application Publication No. 2003 / 0190311. Specific examples include substitutions with leucine at position 251, tyrosine, tryptophan, or phenylalanine at position 252, threonine or serine at position 254, arginine at position 255, glutamine, arginine, serine, threonine, or glutamic acid at position 256, threonine at position 308, proline at position 309, serine at position 311, aspartic acid at position 312, leucine at position 314, arginine, asparagine at position 385, and the like. substitution with threonine or proline at position 386, with arginine or proline at position 387, with proline, asparagine, or serine at position 389, with methionine or threonine at position 428, with tyrosine or phenylalanine at position 434, with histidine, arginine, lysine, or serine at position 433, and / or with histidine, tyrosine, arginine, or threonine at position 436, including any combination thereof. Such modifications optionally increase the affinity of the Fc region for FcRn, thereby increasing the half-life compared to the corresponding, wild-type Fc region.

[0139] Certain variant hybrids or modified Fc regions may have altered solubility compared to the corresponding wild-type Fc sequence. In certain embodiments, such Fc regions may have increased solubility compared to the corresponding wild-type Fc sequence. In other embodiments, variant hybrids or modified Fc regions may have decreased solubility compared to the corresponding wild-type Fc sequence. Solubility can be measured, for example, in vitro (e.g., under physiological conditions) according to routine techniques in the art. Exemplary solubility measurements are described elsewhere herein.

[0140] The variant Fc region can also have one or more mutant hinge regions, for example, as described in U.S. Patent Application Publication No. 2003 / 0118592. For example, one or more cysteines in the hinge region can be deleted or substituted with different amino acids. The mutant hinge region can contain no cysteine ​​residues, or it can contain one, two, or three fewer cysteine ​​residues than the corresponding wild-type hinge region. In some embodiments, Fc regions with this type of mutant hinge region exhibit reduced dimerization ability compared to the wild-type Ig hinge region.

[0141] In certain embodiments, the antibody or antigen-binding fragment thereof is incubated for about or at least about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 12 hours, about 18 hours, about 20 hours, about 24 hours, about 36 hours, about 40 hours, about 50 hours, about 60 hours, about 70 hours, about 80 hours, about 90 hours, about 100 hours, about 120 hours, about 160 hours, about 180 hours, about 240 hours, about 280 hours, about 360 hours, about 380 hours, about 400 hours, about 400 hours, about 500 hours, about 600 hours, about 700 hours, about 800 hours, about 900 hours, about 1000 hours, about 1200 hours, about 1400 hours, about 1600 hours, about 1800 hours, about 2000 hours, about 24 ... A biological half-life of 4 hours, about 30 hours, about 36 hours, about 40 hours, about 48 hours, about 50 hours, about 60 hours, about 70 hours, about 72 hours, about 80 hours, about 84 hours, about 90 hours, about 96 hours, about 120 hours, or about 144 hours or more, or about 1 week, or about 2 weeks, or about 3 weeks, or about 4 weeks, or about 5 weeks, or about 6 weeks or more, or any intervening half-life, including all ranges therebetween.

[0142] In some embodiments, the antibody or antigen-binding fragment thereof has a T of about or at least about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or 75°C. m In some embodiments, the antibody or antigen-binding fragment thereof has a T of about 65° C. or higher, for example, in PBS (phosphate buffered saline). m It has.

[0143] In some embodiments, the antibody or antigen-binding fragment thereof is conjugated to one or more cytotoxic or chemotherapeutic agents. General examples of cytotoxic or chemotherapeutic agents include, but are not limited to, alkylating agents, antimetabolites, anthracyclines, antitumor antibiotics, platinum, type I topoisomerase inhibitors, type II topoisomerase inhibitors, vinca alkaloids, and taxanes. Specific examples of cytotoxic or chemotherapeutic agents include, but are not limited to, cyclophosphamide, cilengitide, lomustine (CCNU), melphalan, procarbazine, carmustine (BCNU), enzastaurin, busulfan, daunorubicin, doxorubicin, gefitinib, and erlotinib. Idarubicin, temozolomide, epirubicin, mitoxantrone, bleomycin, cisplatin, carboplatin, oxaliplatin, camptothecin, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, teniposide, temsirolimus, everolimus, vincristine, vinblastine, vinorelbine, vindesine, CT52923, paclitaxel, imatinib, dasatinib, sorafenib, pazopanib, sunitonib, vatalanib, gefitinib These include futinib, erlotinib, AEE-788, dichloroacetic acid, tamoxifen, fasudil, SB-681323, semaxanib, donepizil, galantamine, memantine, rivastigmine, tacrine, rasigline, naltrexone, lubiprostone, safinamide, istradefylline, pimavanserin, pitolisant, isradipine, pridopidine (ACR16), tetrabenazine, bexarotene, glatirimer acetate, fingolimod, and mitoxantrone, including pharmaceutically acceptable salts and acids thereof. Further examples of cytotoxic or chemotherapeutic agents include alkylating agents, such as thiotepa, cyclophosphamide (CYTOXAN™), and the like; alkylsulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, metholedopa, uredopa, and the like;Ethylenimines and methylameramines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards, such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembrine, phenesterine, prednimustine, trofosfamide, uracil mustard, and the like; nitrothreas, such as carmustine, chlorozotocin, fotem antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, martiromycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, povidone, ribomycin ... tofilomycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, etc.; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU), etc.; folic acid analogs, such as denopterin, methotrexate, pteropterin, trimetrexate, etc.; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, etc.; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmov acetaminophen, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU, etc.; androgens, such as calsterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone, etc.; antiadrenal agents, such as aminoglutethimide, mitotane, trilostane, etc.; folic acid supplements, such as furoic acid, etc.; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestravcil; bisantrene; edatraxate; defoamine; demecolcine; diaziquone; erformitin;Elliptinium acetate; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Lonidamine; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK; Razoxane; Sizofiran; Spirogermanium; Tenuazonic acid; Triazicone; 2,2',2"-Trichlorotriethylamine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"); Cyclophosphamide; Thiotepa; Taxoids, such as paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ) and doxetaxel (TAXOTERE®, Rhône-Poulenc Roller, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; Xeloda; ibandronate; CPT-11; the topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoic acid derivatives such as Targretin™ (bexarotene); Panretin™ (alitretinoin); ONTAK™ (denileukin diftitox); esperamycin; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0144] In some embodiments, the antibodies disclosed herein are conjugated or operably linked to a radioisotope to form radioconjugates and / or macrocyclic chelators useful for complexing radioactive metal ions. A variety of radioisotopes are available for the production of radioconjugated antibodies. Examples include, but are not limited to: 90 Y, 123 I,125 I, 131 I, 186 Re, 188 Re, 211 At, and 212 In certain embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), which can be attached to the antibody via a linker molecule. Such linker molecules are generally known in the art and are described in Denardo et al., 1998, Clin Cancer Res. 4:2483-90; Peterson et al., 1999, Bioconjug. Chem. 10:553; and Zimmerman et al., 1999, Nucl. Med. Biol. 26:943-50.

[0145] Other modifications of the antibodies (and polypeptides) of this disclosure are also contemplated herein. For example, in some embodiments, the antibody is linked to one of a variety of nonproteinaceous polymers, such as polyethylene glycol, polypropylene glycol, polyoxyalkylenes, or copolymers of polyethylene glycol and polypropylene glycol. In some embodiments, the antibody is encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacrylate) microcapsules, respectively), in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980).

[0146] The antibodies or antigen-binding fragments thereof may be used in any of the compositions, methods, and / or kits described herein and may be combined with one or more of the additional agents described herein.

[0147] Methods of Use and Pharmaceutical Compositions Certain embodiments relate to methods of treating, ameliorating symptoms, and / or slowing the progression of a disease or condition in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding fragment thereof that binds to IL-18BP, or a pharmaceutical composition comprising the same, as described herein. Also included are methods of stimulating an immune response, e.g., an IL-18-mediated immune response, in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some examples, the antibody or antigen-binding fragment thereof antagonizes the binding / signaling activity between IL-18BP and its ligand, IL-18, thereby increasing IL-18-mediated signaling or activity (e.g., increased induction of IFN-gamma, CXCL10, and / or TNFα). In some embodiments, the disease or condition is cancer or tumor, or an infectious disease, as described above. In some embodiments, the disease is any disease in which activation of the immune system may be beneficial.

[0148] In some embodiments, the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, such as a lymphoproliferative disorder, a myeloproliferative disorder, proliferative enterocolitis, proliferative diabetic retinopathy, or a proliferative renal disease, as described above. In some examples, the cancer or tumor expresses or overexpresses IL-18BP, IL-18, or both. In some examples, the proliferative disease or disorder is associated with increased expression of IL-18BP, IL-18, or both. In some examples, the cancer is a primary cancer. In some examples, the cancer is a metastatic cancer. Accordingly, certain embodiments include a method of treating, reducing the severity of, or preventing cancer in a patient in need thereof, comprising administering to the patient a composition described herein, wherein the antibody or antigen-binding fragment thereof is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing cancer.

[0149] Examples of cancer include, but are not limited to, bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (HCC), and hepatocellular carcinoma (HCC). Cancers include, for example, alveolar carcinoma (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer. In certain embodiments, the cancer is metastatic cancer, e.g., that has metastasized to bone.

[0150] Also provided are antibodies, antigen-binding fragments thereof, or pharmaceutical compositions of the present disclosure for use as pharmaceuticals. The antibodies, fragments thereof, or pharmaceutical compositions of the present disclosure may be for use in any of the therapeutic methods disclosed herein. In certain embodiments, the antibodies, fragments thereof, or pharmaceutical compositions of the present disclosure may be for use in a method of treating, ameliorating the symptoms of, and / or slowing the progression of any of the diseases or disorders disclosed herein, such as cancer, tumors, or other proliferative diseases or disorders, such as lymphoproliferative disorders, myeloproliferative disorders, proliferative enterocolitis, proliferative diabetic retinopathy, or proliferative kidney diseases.

[0151] Certain embodiments include combination therapies, e.g., the combination therapies include administering a pharmaceutical composition described herein (comprising an anti-IL-18BP antibody or antigen-binding fragment thereof) in combination with one or more additional therapeutic agents, e.g., an immunostimulatory agent, an immune checkpoint modulator, and / or a chemotherapeutic agent. In some embodiments, the additional therapeutic agent comprises IL-18, including human IL-18 (or a functional variant or fragment thereof).

[0152] In some embodiments, the additional therapeutic agent comprises an immune checkpoint modulator. Specific examples of immune checkpoint modulators include "antagonists" of one or more inhibitory immune checkpoint molecules and "agonists" of one or more stimulatory immune checkpoint molecules. Generally, immune checkpoint molecules are components of the immune system that either upregulate (costimulatory molecules) or downregulate (decreasing) signals. Targeting immune checkpoint molecules has therapeutic potential in cancer, as cancer cells can disrupt the natural function of immune checkpoint molecules (see, e.g., Sharma and Allison, Science. 348:56-61, 2015; Topalian et al., Cancer Cell. 27:450-461, 2015; Pardoll, Nature Reviews Cancer. 12:252-264, 2012). In some embodiments, an immune checkpoint modulator (e.g., antagonist, agonist) "binds" or "specifically binds" to one or more immune checkpoint molecules, as described herein.

[0153] In certain embodiments, the immune checkpoint modulator is a polypeptide or peptide. The terms "peptide" and "polypeptide" are used interchangeably herein, although in certain instances, the term "peptide" can refer to a short polypeptide, e.g., a polypeptide consisting of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 amino acids (including all integers and ranges therebetween, e.g., 5-10, 8-12, 10-15). Polypeptides and peptides may be composed of naturally occurring and / or non-naturally occurring amino acids, as described herein.

[0154] Antibodies are also included as polypeptides. Thus, in some embodiments, the immune checkpoint modulating polypeptide agent is an "antibody or antigen-binding fragment thereof," as described herein.

[0155] In some embodiments, the agent is or includes a "ligand," e.g., a native ligand of an immune checkpoint molecule. "Ligand" generally refers to a substance or molecule that forms a complex with a target molecule (e.g., a biomolecule) to achieve a biological purpose, and includes "protein ligands." Protein ligands generally generate a signal by binding to a site on a target molecule or target protein. Thus, certain agents are protein ligands that naturally bind to an immune checkpoint molecule and generate a signal. Also included are "modified ligands." Modified ligands are, for example, pharmacokinetic modifiers, e.g., protein ligands fused to an Fc region derived from an immunoglobulin.

[0156] The binding properties of a polypeptide can be quantified using methods known in the art (see Davies et al., Annual Rev. Biochem. 59:439-473, 1990). In some embodiments, the polypeptide binds to a target molecule, e.g., an immune checkpoint molecule or epitope thereof, with a binding affinity of about ≦10 -7 M ~ about 10 -8 M. In some embodiments, the equilibrium dissociation constant is about ≦10 -9 M~approx.≦10 -10 In certain exemplary embodiments, the polypeptide has an affinity (Kd or EC for a target described herein to which it specifically binds) of about, at least about, or less than about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, or 50 nM. 50 )

[0157] In some embodiments, the agent is a "small molecule," which refers to an organic compound of synthetic or biological origin (biomolecule), but which is often not a polymer. Organic compounds refer to a large class of chemical compounds whose molecules contain carbon, typically excluding those that contain only carbonates, simple oxides of carbon, or cyanides. "Biomolecules" generally refer to organic molecules produced by living organisms, including large macromolecules (biopolymers), such as peptides, polysaccharides, and nucleic acids, as well as small molecules, such as primary and secondary metabolites, lipids, phospholipids, glycolipids, sterols, glycerolipids, vitamins, and hormones. "Polymer" generally refers to a large molecule or macromolecule composed of repeating structural units, typically linked by covalent bonds.

[0158] In certain embodiments, small molecules have a molecular weight of about or less than about 1000-2000 daltons, typically about 300-700 daltons, including less than about or about 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 500, 650, 600, 750, 700, 850, 800, 950, 1000, or 2000 daltons.

[0159] Certain small molecules may have the property of "specific binding" as described for polypeptides herein, e.g., antibodies. For example, in some embodiments, a small molecule binds to a target, e.g., an immune checkpoint molecule, with a binding affinity (Kd or EC) of about, at least about, or less than about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, or 50 nM. 50 ) and binds specifically to it.

[0160] In some embodiments, the immune checkpoint modulator is an antagonist or inhibitor of one or more inhibitory immune checkpoint molecules, including Programmed Death-Ligand 1 (PD-L1), Programmed Death-Ligand 2 (PD-L2), Programmed Death 1 (PD-1), Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA-4), Indoleamine 2,3-Dioxygenase (IDO), Tryptophan 2,3-Dioxygenase (TDO), T-cell Immunoglobulin Domain and Mucin Domain 3 (TIM-3), Lymphocyte Activation Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T Lymphocyte Attenuator (BTLA), CD160, and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0161] In certain embodiments, the agent is a PD-1 (receptor) antagonist or inhibitor, the targeting of which has been shown to restore immune function in the tumor environment (see, e.g., Phillips et al., Int Immunol. 27:39-46, 2015). PD-1 is a cell surface receptor belonging to the immunoglobulin superfamily and expressed on T cells and pro-B cells. PD-1 interacts with two ligands, PD-L1 and PD-L2. PD-1 functions as an inhibitory immune checkpoint molecule, for example, by reducing or preventing T cell activation, which in turn reduces autoimmunity and promotes self-tolerance. The inhibitory effect of PD-1 is achieved, at least in part, through a dual mechanism: promoting apoptosis of antigen-specific T cells in lymph nodes while reducing apoptosis of regulatory T cells (suppressor T cells). Some examples of PD-1 antagonists or inhibitors include antibodies or antigen-binding fragments thereof, or small molecules that specifically bind to PD-1 and reduce one or more of its immunosuppressive activities, such as downstream signaling or interaction with PD-L1. Specific examples of PD-1 antagonists or inhibitors include the antibodies nivolumab, pembrolizumab, PDR001, MK-3475, AMP-224, AMP-514, and pidilizumab, and antigen-binding fragments thereof (see, e.g., U.S. Patent Nos. 8,008,449, 8,993,731, 9,073,994, 9,084,776, 9,102,727, 9,102,728, 9,181,342, 9,217,034, 9,387,247, 9,492,539, 9,492,540, and U.S. Patent Application Nos. 2012 / 0039906 and 2015 / 0203579).

[0162] In some embodiments, the agent is a PD-L1 antagonist or inhibitor. As described above, PD-L1 is one of the natural ligands of the PD-1 receptor. Common examples of PD-L1 antagonists or inhibitors include antibodies or antigen-binding fragments thereof, or small molecules that specifically bind to PD-L1 and reduce one or more of its immunosuppressive activities, such as binding to the PD-1 receptor. Specific examples of PD-L1 antagonists include the antibodies atezolizumab (MPDL3280A), avelumab (MSB0010718C), and durvalumab (MEDI4736), and antigen-binding fragments thereof (see, e.g., U.S. Patent Nos. 9,102,725, 9,393,301, 9,402,899, and 9,439,962).

[0163] In some embodiments, the agent is a PD-L2 antagonist or inhibitor. As discussed above, PD-L2 is one of the natural ligands of the PD-1 receptor. Common examples of PD-L2 antagonists or inhibitors include antibodies or antigen-binding fragments thereof, or small molecules that specifically bind to PD-L2 and reduce one or more of its immunosuppressive activities, such as binding to the PD-1 receptor.

[0164] In some embodiments, the agent is a CTLA-4 antagonist or inhibitor. CTLA4 or CTLA-4 (cytotoxic T-lymphocyte-associated protein 4), also known as CD152 (cluster of differentiation 152), is a protein receptor that functions as an inhibitory immune checkpoint molecule by sending an inhibitory signal to T cells when it binds to, for example, CD80 or CD86 on the surface of antigen-presenting cells. Common examples of CTLA-4 antagonists or inhibitors include antibodies or antigen-binding fragments or small molecules that specifically bind to CTLA-4. Specific examples include the antibodies ipilimumab and tremelimumab, and their antigen-binding fragments. At least part of the activity of ipilimumab is believed to be mediated by antibody-dependent cell-mediated cytotoxicity (ADCC) killing of CTLA-4-expressing suppressor Tregs.

[0165] In some embodiments, the agent is an IDO antagonist or inhibitor, or a TDO antagonist or inhibitor. IDO and TDO are tryptophan catabolism enzymes with immunosuppressive properties. For example, IDO is known to suppress T cells and NK cells, generate and activate Tregs and myeloid-derived suppressor cells, and promote tumor angiogenesis. Common examples of IDO and TDO antagonists or inhibitors include antibodies or antigen-binding fragments, or small molecules that specifically bind to IDO or TDO (see, for example, Platten et al., Front Immunol. 5:673, 2014) and reduce or inhibit one or more immunosuppressive activities. Specific examples of IDO antagonists or inhibitors include indoximod (NLG-8189), 1-methyl-tryptophan (1MT), β-carboline (norharman, 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat (see, e.g., Sheridan, Nature Biotechnology. 33: 321-322, 2015). Specific examples of TDO antagonists or inhibitors include 680C91 and LM10 (see, e.g., Pilotte et al., PNAS USA. 109: 2497-2502, 2012).

[0166] In some embodiments, the agent is a TIM-3 antagonist or inhibitor. T-cell immunoglobulin domain and mucin domain 3 (TIM-3) is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines. TIM-3 also acts as a negative regulator of Th1 / Tc1 function by inducing cell death upon interaction with its ligand, galectin-9. TIM-3 contributes to a suppressive tumor microenvironment, and its overexpression is associated with poor prognosis in various cancers (see, e.g., Li et al., Acta Oncol. 54:1706-13, 2015). Common examples of TIM-3 antagonists or inhibitors include antibodies, antigen-binding fragments thereof, or small molecules that specifically bind to TIM-3 and reduce or inhibit one or more of its immunosuppressive activities.

[0167] In some embodiments, the agent is an antagonist or inhibitor of LAG-3. Lymphocyte Activation Gene-3 (LAG-3) is expressed on activated T cells, natural killer cells, B cells, and plasmacytoid dendritic cells. Like CTLA-4 and PD-1, LAG-3 negatively regulates T cell proliferation, activation, and homeostasis (see, e.g., Workman and Vignali, European Journal of Immun. 33:970-9, 2003; and Workman et al., Journal of Immun. 172:5450-5, 2004) and has been reported to play an important role in the suppressive function of Tregs (see, e.g., Huang et al., Immunity. 21:503-13, 2004). LAG3 also maintains CD8+ T cells in a tolerogenic state and associates with PD-1 to maintain the exhaustion of CD8 T cells. General examples of LAG-3 antagonists or inhibitors include antibodies, antigen-binding fragments thereof, or small molecules that specifically bind to LAG-3 and inhibit one or more of its immunosuppressive activities. Specific examples include the antibody BMS-986016 and antigen-binding fragments thereof.

[0168] In some embodiments, the agent is a VISTA antagonist or inhibitor. V-domain Ig suppressor of T cell activation (VISTA) is an inhibitory immune checkpoint regulator that is expressed primarily on hematopoietic cells, suppresses T cell activation, induces Foxp3 expression, and is highly expressed in the tumor microenvironment. VISTA suppresses anti-tumor T cell responses in the tumor microenvironment (see, e.g., Lines et al., Cancer Res. 74:1924-32, 2014). Common examples of VISTA antagonists or inhibitors include antibodies, antigen-binding fragments thereof, or small molecules that specifically bind to VISTA and reduce one or more of its immunosuppressive activities.

[0169] In some embodiments, the agent is a BTLA antagonist or inhibitor. Expression of B- and T-lymphocyte attenuator (BTLA; CD272) is induced during T cell activation and inhibits T cells through interaction with tumor necrosis family receptors (TNF-R) and the B7 family of cell surface receptors. BTLA is a ligand for tumor necrosis factor (receptor) superfamily, member 14 (TNFRSF14) and is also known as herpes virus entry mediator (HVEM). The BTLA-HVEM complex negatively regulates T cell immune responses, for example, by inhibiting the function of human CD8+ cancer-specific T cells (see, e.g., Derre et al., J Clin Invest 120:157-67, 2009). Typical examples of BTLA antagonists or inhibitors include antibodies, antigen-binding fragments thereof, or small molecules that specifically bind to BTLA-4 and reduce one or more of its immunosuppressive activities.

[0170] In some embodiments, the agent is an antagonist or inhibitor of HVEM, e.g., an antagonist or inhibitor that specifically binds to HVEM and interferes with its interaction with BTLA or CD 160. Common examples of HVEM antagonists or inhibitors include antibodies, or antigen-binding fragments thereof, or small molecules that specifically bind to HVEM and optionally reduce the HVEM / BTLA and / or HVEM / CD 160 interaction, thereby reducing one or more of the immunosuppressive activities of HVEM.

[0171] In some embodiments, the agent is a CD160 antagonist or inhibitor, e.g., an antagonist or inhibitor that specifically binds to CD160 and interferes with its interaction with HVEM. Common examples of CD160 antagonists or inhibitors include antibodies, or antigen-binding fragments thereof, or small molecules that specifically bind to CD160 and optionally reduce the CD160 / HVEM interaction, thereby reducing or inhibiting one or more of its immunosuppressive activities.

[0172] In some embodiments, the agent is a TIGIT antagonist or inhibitor. T cell Ig and ITIM domain (TIGIT) is a co-inhibitory receptor present on the surface of various lymphoid cells that suppresses anti-tumor immunity, for example, via Tregs (Kurtulus et al., J Clin Invest. 125:4053-4062, 2015). Common examples of TIGIT antagonists or inhibitors include antibodies, antigen-binding fragments thereof, or small molecules that specifically bind to TIGIT and reduce one or more of its immunosuppressive activities (see, e.g., Johnston et al., Cancer Cell. 26:923-37, 2014).

[0173] In certain embodiments, the immune checkpoint modulator is an agonist of one or more stimulatory immune checkpoint molecules, examples of which include OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

[0174] In some embodiments, the agent is an OX40 agonist. OX40 (CD134) promotes the proliferation of effector and memory T cells and suppresses the differentiation and activity of T regulatory cells (see, e.g., Croft et al., Immunol Rev. 229:173-91, 2009). Its ligand is OX40L (CD252). OX40 signaling influences both T cell activation and survival, and therefore plays an important role in initiating antitumor immune responses in lymph nodes and maintaining antitumor immune responses in the tumor microenvironment. Common examples of OX40 agonists include antibodies or antigen-binding fragments thereof, or small molecules, or ligands that specifically bind to OX40 and increase one or more of its immunostimulatory activities. Specific examples include OX86, OX-40L, Fc-OX40L, GSK3174998, MEDI0562 (humanized OX40 agonist), MEDI6469 (murine OX4 agonist), and MEDI6383 (OX40 agonist), and antigen-binding fragments thereof.

[0175] In some embodiments, the agent is a CD40 agonist. CD40 is expressed on antigen-presenting cells (APCs) and some malignant tumors. Its ligand is CD40L (CD154). On APCs, ligation results in upregulation of costimulatory molecules, potentially eliminating the need for T cell help in antitumor immune responses. CD40 agonist therapy plays an important role in APC maturation and their migration from tumors to lymph nodes, resulting in improved antigen presentation and T cell activation. Anti-CD40 agonist antibodies have induced robust responses and durable anticancer immunity in animal models, effects mediated, at least in part, by cytotoxic T cells (see, e.g., Johnson et al., Clin Cancer Res. 21:1321-1328, 2015; and Vonderheide and Glennie, Clin Cancer Res. 19:1035-43, 2013). Common examples of CD40 agonists include antibodies or antigen-binding fragments thereof, or small molecules, or ligands that specifically bind to CD40 and increase one or more of its immunostimulatory activities. Specific examples include sotigalilmab, CP-870,893, dacetuzumab, Chi Lob 7 / 4, ADC-1013, CD40L, rhCD40L, and antigen-binding fragments thereof.

[0176] In some embodiments, the agent is a GITR agonist. Glucocorticoid-Induced TNFR family Related gene (GITR) increases T cell expansion, inhibits Treg suppressive activity, and prolongs T effector cell survival. GITR agonists have been shown to promote anti-tumor responses by disrupting the stability of the Treg lineage (see, e.g., Schaer et al., Cancer Immunol Res. 1:320-31, 2013). These diverse mechanisms indicate that GITR plays an important role in initiating immune responses in lymph nodes and maintaining immune responses in tumor tissues. Its ligand is GITRL. Common examples of GITR agonists include antibodies or antigen-binding fragments thereof, or small molecules or ligands that specifically bind to GITR and increase one or more of its immunostimulatory activities. Specific examples include GITRL, INCAGN01876, DTA-1, MEDI1873, and antigen-binding fragments thereof.

[0177] In some embodiments, the agent is a CD137 agonist. CD137 (4-1BB) is a member of the tumor necrosis factor (TNF) receptor family, and cross-linking of CD137 enhances T cell proliferation, IL-2 secretion, survival, and cytolytic activity. CD137-mediated signaling also protects T cells, such as CD8+ T cells, from activation-induced cell death. Common examples of CD137 agonists include antibodies or antigen-binding fragments thereof, small molecules, or ligands that specifically bind to CD137 and increase one or more of its immunostimulatory activities. Specific examples include CD137 (or 4-1BB) ligands (see, e.g., Shao and Schwarz, J. Leukoc. Biol. 89:21-9, 2011), and utomirumab, an antibody containing an antigen-binding fragment thereof.

[0178] In some embodiments, the agent is a CD27 agonist. Stimulation of CD27 increases the antigen-specific expansion of naive T cells and contributes to the long-term maintenance of T cell memory and T cell immunity. Its ligand is CD70. Targeting human CD27 with agonistic antibodies stimulates T cell activation and antitumor immunity (see, e.g., Thomas et al., Oncoimmunology. 2014;3:e27255.doi:10.4161 / onci.27255; and He et al., J Immunol. 191:4174-83, 2013). General examples of CD27 agonists include antibodies or antigen-binding fragments, or small molecules, or ligands that specifically bind to CD27 and increase one or more of its immunostimulatory activities. Specific examples include the antibodies valilumab and CDX-1127 (1F5), which contain CD70 and antigen-binding fragments thereof.

[0179] In some embodiments, the agent is a CD28 agonist. CD28 is constitutively expressed on CD4+ T cells and a subset of CD8+ T cells. Its ligands include CD80 and CD86, and stimulation of these ligands increases T cell proliferation. Common examples of CD28 agonists include antibodies or antigen-binding fragments, or small molecules, or ligands that specifically bind to CD28 and increase one or more of its immunostimulatory activities. Specific examples include CD80, CD86, the antibody TAB08, and antigen-binding fragments thereof.

[0180] In some embodiments, the agent is a CD226 agonist. CD226 shares a ligand with TIGIT and is a stimulatory receptor opposite to TIGIT. CD226 engagement enhances T cell activation (see, for example, Kurtulus et al., J Clin Invest. 125:4053-4062, 2015; Bottino et al., J Exp Med. 1984:557-567, 2003; and Tahara-Hanaoka et al., Int Immunol. 16:533-538, 2004). Common examples of CD226 agonists include antibodies or antigen-binding fragments, or small molecules, or ligands (e.g., CD112, CD155) that specifically bind to CD226 and increase one or more of its immunostimulatory activities.

[0181] In some embodiments, the agent is an HVEM agonist. Herpesvirus entry mediator (HVEM), also known as tumor necrosis factor receptor superfamily member 14 (TNFRSF14), is a human cell surface receptor of the TNF receptor superfamily. HVEM is present on a variety of cells, including T cells, APCs, and other immune cells. Unlike other receptors, HVEM is expressed at high levels on resting T cells and is downregulated upon activation. HVEM signaling has been shown to play an important role in the early stages of T cell activation and during the expansion of tumor-specific lymphocyte populations in lymph nodes. Common examples of HVEM agonists include antibodies or antigen-binding fragments, small molecules, or ligands that specifically bind to HVEM and increase one or more of its immunostimulatory activities.

[0182] In some embodiments, the additional therapeutic agent comprises a chemotherapeutic agent, e.g., a small molecule chemotherapeutic agent, non-limiting examples of which include alkylating agents, antimetabolites, cytotoxic antibiotics, topoisomerase inhibitors (type 1 or type II), and anti-microtubule agents, among others.

[0183] Examples of alkylating agents include nitrogen mustards (e.g., mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosoureas (e.g., N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, and streptozotocin), tetrazines (e.g., dacarbazine, mitozolomide, and temozolomide), aziridines (e.g., thiotepa, mitomycin, and diaziquone (AZQ)), cisplatin and its derivatives (e.g., carboplatin and oxaplatin), and non-classical alkylating agents (optionally procarbazine and hexamethylmelamine).

[0184] Examples of antimetabolites include antifolates (e.g., methotrexate and pemetrexed), fluoropyrimidines (e.g., 5-fluorouracil and capecitabine), deoxynucleoside analogs (e.g., ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (e.g., thioguanine and mercaptopurine).

[0185] Examples of cytotoxic antibiotics include anthracyclines (e.g., doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, and mitoxantrone), bleomycin, mitomycin C, mitoxantrone, and actinomycin. Examples of topoisomerase inhibitors include camptothecin, irinotecan, topotecan, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, mervalone, and aclarubicin.

[0186] Examples of anti-microtubule agents include taxanes (eg, paclitaxel and docetaxel), and vinca alkaloids (eg, vinblastine, vincristine, vindesine, vinorelbine).

[0187] In certain embodiments, the methods and compositions described herein are sufficient to cause a statistically significant decrease in the amount of viable tumor, e.g., at least a 10%, 20%, 30%, 40%, 50% or greater decrease in tumor mass, or tumor regression as indicated by an altered (e.g., statistically significantly decreased) scan dimension. In some embodiments, the methods and compositions described herein reduce the rate of cancer growth (e.g., in vivo or in vitro, including cancer cells isolated from a biopsy or other sample and grown in vitro) by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to an untreated control. In some examples, the methods and compositions described herein reduce cancer cell initiation, migration, adhesion, invasiveness, and / or metastasis by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to untreated controls. In some examples, the methods and compositions described herein reduce angiogenesis in the tumor environment by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to untreated controls.

[0188] In certain embodiments, the disease or condition is myelodysplastic syndrome (MDS) (see, e.g., Wang et al., Blood. 140(Supplement 1):12297, 2022). For example, in response, methods of antagonizing IL-18BP are a viable approach. MDS refers to a group of cancers in which immature blood cells in the bone marrow fail to mature and, as a result, fail to develop into healthy blood cells. Accordingly, certain embodiments include methods of treating, reducing the severity of, or preventing MDS in a patient in need thereof, comprising administering to the patient a composition described herein, wherein the antibody or antigen-binding fragment thereof is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing MDS.

[0189] In some embodiments, the disease or condition is an infectious disease. For example, in certain embodiments, the infectious disease is selected from a viral (see, e.g., Vecchie et al., J Cell Physiol. 236(3):1638-1657, 2021), bacterial (see, e.g., Kinoshita et al., Ann Surg. 240(2):313-20, 2004), fungal (e.g., yeast), and protozoan infection. Accordingly, some embodiments include a method of treating, reducing the severity of, or preventing an infectious disease in a patient in need thereof, the method comprising administering to the patient a composition described herein, wherein the antibody or antigen-binding fragment thereof is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing the infectious disease.

[0190] In some embodiments, the methods and compositions described herein increase the median survival time of a subject by 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 15 weeks, 20 weeks, 25 weeks, 30 weeks, 40 weeks, or more. In certain embodiments, the methods and compositions described herein increase the median survival time of a subject by 1 year, 2 years, 3 years, or more. In some embodiments, the methods and compositions described herein increase progression-free survival by 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, or more. In certain embodiments, the methods and compositions described herein increase progression-free survival by 1 year, 2 years, 3 years, or more.

[0191] In certain embodiments, the methods and compositions described herein are sufficient to result in stable disease, hi certain embodiments, the methods and compositions described herein are sufficient to result in a clinically relevant reduction in symptoms of a particular disease indication known to a skilled clinician.

[0192] For in vivo use, certain embodiments include pharmaceutical compositions comprising an antibody or antigen-binding fragment thereof described herein and a pharmaceutically acceptable carrier. To prepare a therapeutic or pharmaceutical composition, an effective or desired amount of one or more agents is mixed with any pharmaceutical carrier or excipient known to those of skill in the art to be appropriate for the particular agent and / or mode of administration. The pharmaceutical carrier may be liquid, semi-liquid, or solid. Solutions or suspensions used for parenteral, intradermal, intraocular, subcutaneous, direct instillation into the bladder, or topical application may contain, for example, sterile diluents (such as water), saline (e.g., phosphate-buffered saline; PBS), fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents (such as benzyl alcohol and methylparabens); antioxidants (such as ascorbic acid and sodium bisulfite) and chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); buffers (such as acetates, citrates, and phosphates). If administered intravenously (e.g., by IV infusion), suitable carriers include saline or phosphate buffered saline (PBS), as well as solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0193] Administration of the agents described herein, in pure form or in suitable therapeutic or pharmaceutical compositions, can be via any of the accepted modes of administration for agents that perform similar functions. Therapeutic or pharmaceutical compositions can be prepared by combining the agent-containing composition with a suitable physiologically acceptable carrier, diluent, or excipient, and can be formulated into preparations in solid, semi-solid, liquid, or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Other pharmaceutically active ingredients (including other small molecules described elsewhere herein) and / or suitable excipients, such as salts, buffers, and stabilizers, can also be present in the composition, but need not be.

[0194] Administration can be achieved by a variety of different routes, including oral, parenteral, nasal, intravenous, intraocular, intradermal, intramuscular, subcutaneous, placement in the bladder, or topical. The preferred mode of administration depends on the nature of the condition being treated or prevented. Certain embodiments include administration by IV infusion.

[0195] Carriers can include, for example, pharmaceutically or physiologically acceptable carriers, excipients, or stabilizers that are nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. Often, the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include buffers, such as phosphate, citrate, other organic acids, and the like; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, histidine, and / or lysine, and the like; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or non-ionic surfactants, such as polysorbate 20 (TWEEN™), polyethylene glycol (PEG), poloxamer (PLURONICS™), and the like.

[0196] In some embodiments, one or more agents can be encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacrylate) microcapsules, respectively), in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980). The particles or liposomes may further comprise other therapeutic or diagnostic agents.

[0197] The exact dosage and duration of treatment are a function of the disease being treated and can be determined empirically using known test protocols or by testing the composition in a model system known in the art and extrapolating from there.Controlled clinical trials can also be carried out.Dosage can also vary depending on the severity of the condition to be alleviated.Pharmaceutical compositions are generally formulated and administered to exert therapeutically useful effects while minimizing undesirable side effects.The composition can be administered once or divided into multiple smaller doses that are administered at regular intervals.For any particular subject, the specific dosage regimen can be adjusted over time according to individual needs.

[0198] Typical routes of administration of these and related therapeutic or pharmaceutical compositions thus include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, ocular, rectal, vaginal, and intranasal. As used herein, the term "parenteral" includes subcutaneous injection, intravenous, intravesical, intramuscular, intrasternal injection, or infusion techniques. Therapeutic or pharmaceutical compositions according to certain embodiments of the present disclosure are formulated so that the active ingredients contained therein are bioavailable upon administration of the composition to a subject or patient. The composition administered to a subject or patient may take the form of one or more dosage units, where, for example, a tablet may be a single dosage unit and a container of a drug described herein in aerosol form may hold multiple dosage units. Actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The compositions to be administered typically contain a therapeutically effective amount of an agent described herein for treatment of the disease or condition of interest.

[0199] Therapeutic or pharmaceutical compositions can be in solid or liquid form.In some embodiments, the carrier is particulate, and the composition is, for example, in tablet or powder form.The carrier can be liquid, and the composition is, for example, oral oil, injectable liquid, or aerosol, which is useful, for example, in inhalation administration.When intended for oral administration, pharmaceutical compositions are preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension, and gel form are included in the form considered herein as either solid or liquid.Certain embodiments include sterile injection solutions.

[0200] As a solid composition for oral administration, the pharmaceutical composition may be formulated into a powder, granules, gel, compressed tablet, pill, capsule, chewing gum, wafer, or the like. Such solid compositions typically contain one or more inert diluents or edible carriers. One or more of the following may also be present: binders such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth, or gelatin; excipients such as starch, lactose, or dextrin; disintegrants such as alginic acid, sodium alginate, Primogel, corn starch, and the like; lubricants such as magnesium stearate or Stereotex; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavorings such as peppermint, methyl salicylate, or orange flavor; and coloring agents. When the pharmaceutical composition is in the form of a capsule, e.g., a gelatin capsule, it may contain, in addition to the above materials, a liquid carrier such as polyethylene glycol or oil.

[0201] Therapeutic or pharmaceutical compositions may be in the form of a liquid, such as an elixir, syrup, solution, gel, emulsion, or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred compositions contain, in addition to the compound, one or more of a sweetener, a preservative, a dye / colorant, and a flavor enhancer. In compositions intended to be administered by injection, one or more of a surfactant, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, and an isotonic agent may be included.

[0202] Liquid therapeutic or pharmaceutical compositions, whether in solution, suspension, or other similar form, may contain one or more of the following adjuvants: sterile diluents, such as water for injection, saline, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils, such as synthetic mono- or diglycerides, polyethylene glycols, glycerin, propylene glycol, or other solvents that serve as solvents or suspending media; antibacterial agents, such as benzyl alcohol or methylparabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, phosphates, and agents for adjusting tonicity, such as sodium chloride or glucose. Parenteral preparations can be enclosed in ampoules, disposable syringes, or multiple-dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. Pharmaceutical compositions for injection are preferably sterilized.

[0203] Liquid therapeutic or pharmaceutical compositions intended for either parenteral, ocular, or oral administration should contain an amount of agent such that an appropriate dosage will be obtained. Typically, this amount is at least 0.01% of the agent of interest in the composition. When intended for oral administration, this amount can vary to be between 0.1 and about 70% by weight of the composition. Certain oral therapeutic or pharmaceutical compositions contain between about 4% and about 75% of the agent of interest. In certain embodiments, therapeutic or pharmaceutical compositions and preparations are prepared so that a parenteral dosage unit contains between 0.01 and 10% by weight of the agent of interest before dilution.

[0204] Therapeutic or pharmaceutical compositions may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment, or gel base. The base may comprise, for example, one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickeners may also be present in therapeutic or pharmaceutical compositions for topical administration. If intended for transdermal administration, the composition may comprise a transdermal patch or iontophoresis device.

[0205] Therapeutic or pharmaceutical compositions may be intended for rectal administration, for example, in the form of suppositories that dissolve in the rectum and release the drug.Compositions for rectal administration may contain an oily base as a suitable non-irritating excipient.Such bases include, but are not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0206] Therapeutic or pharmaceutical compositions may contain various materials that modify the physical form of a solid or liquid dosage unit. For example, the composition may contain a material that forms a coating shell around the active ingredient. The material that forms the coating shell is typically inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in a gelatin capsule. Therapeutic or pharmaceutical compositions in solid or liquid form may contain a component that binds to the drug and thereby assists in the delivery of the compound. Suitable components that can act in this capacity include monoclonal or polyclonal antibodies, one or more proteins, or liposomes.

[0207] Therapeutic or pharmaceutical compositions may consist essentially of dosage units that can be administered as an aerosol. The term "aerosol" is used to refer to a variety of systems, ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by liquefied or compressed gas or by an appropriate pump system that dispenses the active ingredient. The aerosol may be delivered in a single-phase, two-phase, or three-phase system to deliver the active ingredient. Aerosol delivery includes the necessary containers, activators, valves, subcontainers, and the like, which may together form a kit. One skilled in the art can determine a preferred aerosol without undue experimentation.

[0208] The compositions described herein may be prepared with carriers that protect the agent against rapid elimination from the body, such as slow release formulations or coatings. Such carriers include controlled release formulations, including, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid, and others known to those skilled in the art.

[0209] Pharmaceutical compositions can be prepared by methods well known in the pharmaceutical arts. For example, therapeutic or pharmaceutical compositions intended to be administered by injection may contain one or more salts, buffers and / or stabilizers, along with sterile distilled water to form a solution. Surfactants may be added to promote the formation of a homogeneous solution or suspension. Surfactants are compounds that interact non-covalently with drugs to promote the dissolution or homogeneous suspension of the drug in an aqueous delivery system.

[0210] Therapeutic or pharmaceutical compositions may be administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the particular compound used; the metabolic stability and duration of action of the compound; the age, weight, general health, sex, and diet of the subject; the mode and time of administration; the excretion rate; the drug combination; the severity of the particular disorder or condition; and the subject being treated. In some examples, a therapeutically effective daily dose is from about 0.001 mg / kg (i.e., 0.07 mg) to about 100 mg / kg (i.e., 7.0 g) (for a mammal weighing 70 kg); preferably, a therapeutically effective dose is from about 0.01 mg / kg (i.e., 0.7 mg) to about 50 mg / kg (i.e., 3.5 g) (for a mammal weighing 70 kg); more preferably, a therapeutically effective amount is from about 1 mg / kg (i.e., 70 mg) to about 25 mg / kg (i.e., 1.75 g) (for a mammal weighing 70 kg). In some embodiments, the therapeutically effective dose is administered weekly, biweekly, or monthly. In certain embodiments, the therapeutically effective dose is administered weekly, biweekly, or monthly, e.g., at a dose of about 1-10 or 1-5 mg / kg, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg.

[0211] Also included are patient care kits comprising (a) an antibody or antigen-binding fragment thereof that binds to IL-18BP, as described herein; and, optionally, (b) at least one additional therapeutic agent. In certain kits, (a) and (b) are in separate therapeutic compositions. In some kits, (a) and (b) are in the same therapeutic composition.

[0212] The kits herein may also include one or more additional therapeutic agents or other components appropriate or desirable for the indication being treated or for the desired diagnostic application. The kits herein may also include one or more syringes or other components necessary or desirable to facilitate the intended mode of delivery (e.g., stents, implantable depots, etc.).

[0213] In some embodiments, the patient care kit contains separate containers, dividers, or compartments for the composition and informational material. For example, the composition may be contained in a bottle, vial, or syringe, and the informational material can be included in association with the container. In some embodiments, the separate elements of the kit are contained within a single, undivided container. For example, the composition is contained in a bottle, vial, or syringe having informational material in the form of a label attached thereto. In some embodiments, the kit contains multiple (e.g., packs) individual containers, each containing one or more unit dosage forms of the antibody (e.g., dosage forms described herein) and, optionally, at least one additional therapeutic agent. For example, the kit contains multiple syringes, ampoules, foil packets, or blister packs, each containing a single unit dose of the antibody and, optionally, at least one additional therapeutic agent. The containers of the kit can be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-tight.

[0214] The patient care kit optionally includes a device suitable for administering the composition, such as a syringe, inhaler, dropper (e.g., eye dropper), swab (e.g., cotton swab or wooden swab), or any such delivery device. In some embodiments, the device is an implantable device that dispenses a metered dose of the agent. Also included are methods of providing the kit, for example, by combining the components described herein.

[0215] Expression and purification system Certain embodiments include methods and related compositions for expressing and purifying the anti-IL-18BP antibodies or antigen-binding fragments thereof described herein. Such recombinant anti-IL-18BP antibodies can be conveniently prepared using standard protocols described, for example, in Sambrook, et al. (1989, supra), especially sections 16 and 17; Ausubel et al. (1994, supra), especially chapters 10 and 16; and Coligan et al., Current Protocols in Protein Science (John Wiley & Sons, Inc. 1995-1997), especially chapters 1, 5, and 6. As one general example, an anti-IL-18BP antibody can be prepared by a procedure comprising one or more of the following steps: (a) preparing a construct comprising a polynucleotide sequence encoding an anti-IL-18BP antibody heavy and / or light chain and operably linked to regulatory elements; (b) introducing the construct into a host cell; (c) culturing the host cell to express the anti-IL-18BP antibody; and (d) isolating the anti-IL-18BP from the host cell.

[0216] Specific embodiments thus include polynucleotides encoding the anti-IL-18BP antibodies or antigen-binding fragments thereof described herein, vectors containing the polynucleotides, and host cells containing the polynucleotides and / or vectors. To express the desired polypeptide, a nucleotide sequence encoding an anti-IL-18BP, or a functional equivalent, may be inserted into a suitable expression vector, i.e., a vector containing the necessary elements for transcription and translation of the inserted coding sequence. Methods well known to those skilled in the art may be used to construct expression vectors containing a sequence encoding a polypeptide of interest and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described in Sambrook et al., Molecular Cloning, A Laboratory Manual (1989), and Ausubel et al., Current Protocols in Molecular Biology (1989).

[0217] A variety of expression vector / host systems are known and can be utilized to contain and express polynucleotide sequences, including, but not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems, including mammalian cells, more particularly human cell systems.

[0218] "Control elements" or "regulatory sequences" present in an expression vector are vector enhancers, promoters, and those untranslated regions of the 5' and 3' untranslated regions that interact with host cell proteins to effect transcription and translation. Such elements vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, can be used. For example, when cloning in bacterial systems, inducible promoters can be used, such as the hybrid lacZ promoter of the pBLUESCRIPT phagemid (Stratagene, La Jolla, CA) or the PSPORT1 plasmid (Gibco BRL, Gaithersburg, MD). In mammalian cell systems, promoters from mammalian genes or mammalian viruses are generally preferred. If it is necessary to generate cell lines containing multiple copies of a polypeptide-encoding sequence, SV40- or EBV-based vectors can be advantageously used with appropriate selectable markers.

[0219] In bacterial systems, numerous expression vectors can be selected depending on the intended use of the expressed polypeptide. For example, if large quantities are needed, vectors that direct high-level expression of easily purified fusion proteins can be used. Such vectors include, but are not limited to, multifunctional E. coli cloning and expression vectors, such as BLUESCRIPT (Stratagene), in which the sequence encoding the polypeptide of interest can be ligated in frame with the sequence for the amino-terminal Met and the following 7 residues of β-galactosidase, resulting in the production of a hybrid protein; pIN vectors (Van Heeke & Schuster, J. Biol. Chem. 264:5503-5509 (1989)); and the like. pGEX vectors (Promega, Madison, Wis.) can also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). Generally, such fusion proteins are soluble and can be easily purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins produced in such systems may be engineered to contain heparin, thrombin, or factor XA protease cleavage sites, allowing the cloned polypeptide of interest to be optionally released from the GST moiety.

[0220] Certain embodiments may use E. coli-based expression systems (see, e.g., Structural Genomics Consortium et al., Nature Methods. 5:135-146, 2008). These and related embodiments may rely partially or completely on ligation-independent cloning (LIC) to generate suitable expression vectors. In certain embodiments, protein expression may be controlled by T7 RNA polymerase (e.g., the pET vector series). These and related embodiments may utilize the expression host strain BL21(DE3), a λDE3 lysogen of BL21 that supports T7-mediated expression and is deficient in lon and ompT proteases for improved target protein stability. Also included are expression host strains that carry tRNA-encoding plasmids, which are rarely used in E. coli, such as the ROSETTA™(DE3) and Rosetta 2(DE3) strains. Cell lysis and sample handling can also be improved using reagents sold under the trademarks BENZONASE® nuclease and BUGBUSTER® protein extraction reagent. For cell culture, autoinduction media can improve the efficiency of many expression systems, including high-throughput expression systems. This type of media (e.g., the OVERNIGHT EXPRESS™ autoinduction system) gradually initiates protein expression through a metabolic shift without the addition of artificial inducers, such as IPTG. Certain embodiments use hexahistidine tags (such as those sold under the trademark HIS·TAG® fusions), followed by immobilized metal affinity chromatography (IMAC) purification or related techniques. In certain aspects, however, clinical-grade proteins can be isolated from E. coli inclusion bodies with or without the use of affinity tags (see, e.g., Shimp et al., Protein Expr Purif. 50:58-67, 2006). As a further example, certain embodiments may use a cold-shock inducible E. coli high-yield production system.This is because overexpression of proteins in E. coli at low temperatures improves their solubility and stability (see, e.g., Qing et al., Nature Biotechnology. 22:877-882, 2004).

[0221] Also included are high-density bacterial fermentation systems, such as high-cell-density cultures of Ralstonia eutropha, which allow protein production at cell densities exceeding 150 g / L and expression of recombinant proteins at titers exceeding 10 g / L.

[0222] In the yeast Saccharomyces cerevisiae, numerous vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PHH, can be used. For reviews, see Ausubel et al. (supra) and Grant et al., Methods Enzymol. 153:516-544 (1987). Also included is the Pichia pandoris expression system (see, e.g., Li et al., Nature Biotechnology. 24, 210-215, 2006; and Hamilton et al., Science, 301:1244, 2003). Certain embodiments include yeast systems engineered to selectively glycosylate proteins, including, inter alia, yeast with humanized N-glycosylation pathways (see, e.g., Hamilton et al., Science. 313:1441-1443, 2006; Wildt et al., Nature Reviews Microbiol. 3:119-28, 2005; and Gerngross et al., Nature-Biotechnology. 22:1409-1414, 2004; U.S. Patent Nos. 7,629,163; 7,326,681; and 7,029,872). By way of example only, recombinant yeast cultures can be grown in Fernbach flasks or 15 L, 50 L, 100 L, and 200 L fermentors, among others.

[0223] When a plant expression vector is used, expression of the polypeptide-encoding sequence can be driven by any of a number of promoters. For example, viral promoters, such as the 35S and 19S promoters of CaMV, can be used alone or in combination with the omega leader sequence from TMV (Takamatsu, EMBO J. 6:307-311 (1987)). Alternatively, plant promoters, such as the RUBISCO or small subunit heat shock promoters, can be used (Coruzzi et al., EMBO J. 3:1671-1680 (1984); Broglie et al., Science 224:838-843 (1984); and Winter et al., Results Probl. Cell Differ. 17:85-105 (1991)). These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. Such techniques are described in many publicly available reviews (see, eg, Hobbs in McGraw Hill, Yearbook of Science and Technology, pp. 191-196 (1992)).

[0224] Insect systems can also be used to express polypeptides of interest. For example, in one such system, Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes in Spodoptera frugiperda or Trichoplusia cells. A polypeptide-encoding sequence may be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under control of the polyhedrin promoter. Successful insertion of the polypeptide-encoding sequence renders the polyhedrin gene inactive and produces recombinant virus lacking coat protein. The recombinant virus can then be used to infect, for example, S. frugiperda or Trichoplusia cells in which the polypeptide of interest can be expressed (Engelhard et al., Proc. Natl. Acad. Sci. USA 91:3224-3227 (1994)). Also included are baculovirus expression systems, including those utilizing SF9, SF21, and Tni cells (see, e.g., Murphy and Piwnica-Worms, Curr Protoc Protein Sci. Chapter 5:Unit 5.4, 2001). Insect systems can provide post-translational modifications similar to mammalian systems.

[0225] Many virus-based expression systems are commonly available for mammalian host cells.For example, when adenovirus is used as an expression vector, the sequence encoding the target polypeptide can be ligated into the adenovirus transcription / translation complex consisting of a late promoter and a tripartite leader sequence.Insertion into the non-essential E1 or E3 region of the viral genome can also be used to obtain a viable virus capable of expressing polypeptides in infected host cells (Logan & Shenk, Proc. Natl. Acad. Sci. USA 81:3655-3659 (1984)).Transcription enhancers, such as Rous sarcoma virus (RSV) enhancers, can also be used to increase expression in mammalian host cells.

[0226] Examples of useful mammalian host cell lines are the SV40-transformed monkey kidney CV1 line (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat hepatocytes (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TR1 cells (Mather et al., Annals NYAcad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and human hepatoma line (Hep G2). Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., PNAS USA 77:4216 (1980)); and myeloma cell lines, such as NSO and Sp2 / 0. For a review of suitable specific mammalian host cell lines for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 255-268. Certain preferred mammalian cell expression systems include CHO and HEK293-cell based expression systems.Mammalian expression systems can utilize adherent cell lines in, for example, T-flasks, roller bottles, or cell factories, or suspension cultures, such as 1 L and 5 L spinners, 5 L, 14 L, 40 L, 100 L, and 200 L stirred tank bioreactors, or 20 / 50 L and 100 / 200 L WAVE bioreactors, among others known in the art.

[0227] Also included is cell-free expression of proteins. These and related embodiments typically utilize purified RNA polymerase, ribosomes, tRNA, and ribonucleotides; these reagents may be produced by extraction from cells or from cell-based expression systems.

[0228] Specific initiation signals can also be used to achieve more efficient translation of sequences encoding a polypeptide of interest. Such signals include the ATG initiation codon and adjacent sequences. When a polypeptide-encoding sequence, its initiation codon, and upstream sequences are inserted into an appropriate expression vector, additional transcriptional or translational control signals may not be required. However, when only a coding sequence, or a portion thereof, is inserted, exogenous translational control signals, including the ATG initiation codon, should be provided. Furthermore, the initiation codon should be in the correct reading frame to ensure translation of the entire insert. Exogenous translational elements and initiation codons can be of various origins, both natural and synthetic. Expression efficiency can be enhanced by the inclusion of enhancers appropriate for the particular cell system used, such as those described in the literature (Scharf et al., Results Probl. Cell Differ. 20:125-162 (1994)).

[0229] A host cell line may also be selected for its ability to regulate the expression of inserted sequences or process the expressed protein in the desired manner. Such modifications of polypeptides include, but are not limited to, post-translational modifications, such as acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing, which cleaves a "prepro" form of the protein, may be used to facilitate correct insertion, folding, and / or function. Different host cells, such as yeast, CHO, HeLa, MDCK, HEK293, and W138, as well as bacterial cells that possess or lack specific cellular and characteristic machinery for such post-translational activities, may be selected to ensure the correct modification and processing of the foreign protein.

[0230] For long-term, high-yield production of recombinant proteins, stable expression is generally preferred. For example, cell lines stably expressing a polynucleotide of interest can be transformed using an expression vector, which may contain a viral origin of replication and / or endogenous expression elements, as well as a selectable marker gene on the same or a separate vector. After introduction of the vector, the cells may be grown in an enriched medium for approximately 1-2 days before switching them to a selective medium. The purpose of the selectable marker is to confer resistance to selection; its presence allows the growth and recovery of cells that successfully express the introduced sequence. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate for the cell type. Transient production, such as by transient transfection or infection, can also be used. Exemplary mammalian expression systems suitable for transient production include HEK293 and CHO-based systems.

[0231] Any number of selection systems may be used to recover transformed or transduced cell lines, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223-232 (1977)) and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817-823 (1990)) genes, which can be used in tk- or aprt- cells, respectively. Antimetabolite, antibiotic, or herbicide resistance can also be used as the basis for selection; for example, dhfr, which confers resistance to methotrexate (Wigler et al., PNAS USA. 77:3567-70 (1980)); npt, which confers resistance to aminoglycosides, neomycin, and G-418 (Colbere-Garapin et al., J. Mol. Biol. 150:1-14 (1981)); and als or pat, which confers resistance to chlorsulfuron and phosphinothricin acetyltransferase, respectively (Murry, supra). Additional selectable genes have been described, such as trpB, which allows cells to utilize indole instead of tryptophan, or hisD, which allows cells to utilize histinol instead of histidine (Hartman & Mulligan, Proc. Natl. Acad. Sci. USA 85:8047-51 (1988)). The use of visible markers has gained popularity, with markers such as green fluorescent protein (GFP) and other fluorescent proteins (e.g., RFP, YFP), anthocyanins, β-glucuronidase and its substrate GAS, and luciferase and its substrate luciferin being widely used not only to identify transformants but also to quantify the amount of transient or stable protein expression resulting from a particular vector system (see, e.g., Rhodes et al., Methods Mol. Biol. 55:121-131 (1995)).

[0232] Also included are high-throughput protein production systems, or microproduction systems. Certain embodiments may utilize hexahistidine fusion tags for protein expression and purification on metal chelate-modified slide surfaces or MagneHis Ni-particles (see, e.g., Kwon et al., BMC Biotechnol. 9:72, 2009; and Lin et al., Methods Mol Biol. 498:129-41, 2009). Also included are high-throughput cell-free protein expression systems (see, e.g., S Sitaraman et al., Methods Mol Biol. 498:229-44, 2009). These and related embodiments can be used, for example, to generate antibody microarrays, which can then be used to screen libraries to identify antibodies and antigen-binding domains that interact with the IL-18BP polypeptide of interest.

[0233] A variety of protocols for detecting and measuring the expression of polynucleotide-encoded products using binding agents or antibodies, such as polyclonal or monoclonal antibodies specific for the product, are known in the art. Examples include enzyme-linked immunosorbent assay (ELISA), Western immunoblot, radioimmunoassay (RIA), and fluorescence-activated cell sorting (FACS). These and other assays are described, inter alia, in Hampton et al., Serological Methods, a Laboratory Manual (1990) and Maddox et al., J. Exp. Med. 158:1211-1216 (1983).

[0234] A wide variety of labeling and conjugation techniques are known to those skilled in the art and can be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting polynucleotide-related sequences include oligolabeling, nick translation, end-labeling, or PCR amplification using labeled nucleotides. Alternatively, the sequence, or any portion thereof, can be cloned into a vector for the production of mRNA probes. Such vectors are known in the art and commercially available, and can be used to synthesize RNA probes in vitro by adding a suitable RNA polymerase, such as TT7, T3, or SP6, and labeled nucleotides. These procedures can be performed using various commercially available kits. Suitable reporter molecules or labels that can be used include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents, as well as substrates, cofactors, inhibitors, magnetic particles, and the like.

[0235] Host cells transformed with a polynucleotide sequence of interest may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. Certain embodiments utilize serum-free cell expression systems. Examples include HEK293 cells and CHO cells, which can be grown on serum-free medium (see, e.g., Rosser et al., Protein Expr. Purif. 40:237-43, 2005; and U.S. Patent No. 6,210,922).

[0236] Antibodies, or antigen-binding fragments thereof, produced by recombinant cells may be secreted or contained intracellularly, depending on the sequence and / or vector used. As will be understood by those skilled in the art, expression vectors containing polynucleotides may be designed to contain signal sequences that direct secretion of the encoded polypeptide through a prokaryotic or eukaryotic cell membrane. Other recombinant constructs may be used to link a sequence encoding a polypeptide of interest to a nucleotide sequence encoding a polypeptide domain that facilitates purification and / or detection of soluble proteins. Examples of such domains include cleavable and non-cleavable affinity purification tags and epitope tags, such as avidin, FLAG tags, polyhistidine tags (e.g., 6xHis), cMyc tags, V5 tags, glutathione S-transferase (GST) tags, and others.

[0237] Proteins produced by recombinant cells can be purified and characterized according to a variety of techniques known in the art. Exemplary systems for performing protein purification and analyzing protein purity include fast protein liquid chromatography (FPLC) (e.g., AKTA and Bio-Rad FPLC systems), high-pressure liquid chromatography (HPLC) (e.g., Beckman and Waters HPLC). Exemplary chemistries for purification include ion exchange chromatography (e.g., Q, S), size exclusion chromatography, salt gradients, affinity purification (e.g., Ni, Co, FLAG, maltose, glutathione, protein A / G), gel filtration, reversed-phase, ceramic HYPERD® ion exchange chromatography, and hydrophobic interaction columns (HIC), among others, as known in the art. Also included are analytical methods, such as SDS-PAGE (e.g., Coomassie, silver staining), immunoblot, Bradford, and ELISA, which can typically be utilized during any step of the production or purification process to measure the purity of a protein composition.

[0238] Also included are methods for concentrating anti-IL-18BP antibodies and antigen-binding fragments thereof, and compositions comprising the concentrated soluble protein. In certain embodiments, concentrated solutions of anti-IL-18BP antibodies comprise proteins at concentrations of about 5 mg / mL; or about 8 mg / mL; or about 10 mg / mL; or about 15 mg / mL; or about 20 mg / mL or more.

[0239] In some embodiments, the composition is substantially monodisperse, e.g., wherein the anti-IL-18BP antibody is present predominantly (i.e., at least about 90% or more) in one apparent molecular weight form, as assessed, e.g., by size exclusion chromatography, dynamic light scattering, and / or analytical ultracentrifugation.

[0240] In some aspects, the compositions have at least about 90%, or in some aspects at least about 95% purity, or in some embodiments at least about 98% purity (protein basis). Purity can be determined via any routine analytical method known in the art.

[0241] In some embodiments, the composition has a high molecular weight aggregate content of less than about 10%, less than about 5%, less than about 3%, or less than about 1%. High molecular weight aggregate content can be determined by various analytical techniques, including, for example, size exclusion chromatography, dynamic light scattering, and / or analytical ultracentrifugation.

[0242] Examples of concentration approaches contemplated herein include lyophilization, which is typically used when the solution contains few soluble components other than the protein of interest. Lyophilization is often performed after an HPLC run and can remove most or all volatile components from a mixture. Also included is ultrafiltration, a technique that typically uses one or more selectively permeable membranes to concentrate a protein solution. The membrane allows water and small molecules to pass through and retain the protein; the solution can be forced against the membrane by mechanical pumps, gas pressure, or centrifugation, among other techniques.

[0243] In certain embodiments, anti-IL-18BP antibodies, reagents, or related agents have a purity of at least about 90% as measured according to routine techniques in the art. In certain embodiments, anti-IL-18BP compositions have a purity of at least about 95%. In certain embodiments, such as therapeutic or pharmaceutical compositions, anti-IL-18BP antibody compositions have a purity of at least about 97%, 98%, or 99%. In some embodiments, such as when used as a reference or research reagent, anti-IL-18BP antibodies may be less pure and may have a purity of at least about 50%, 60%, 70%, or 80%. Purity can be measured overall or in relation to selected components, such as other proteins, e.g., purity on a protein basis.

[0244] Purified antibodies can also be characterized according to their biological characteristics. Binding affinity and binding kinetics can be measured according to various techniques known in the art, such as Biacore® and related techniques utilizing surface plasmon resonance (SPR), an optical phenomenon that allows for the detection of unlabeled interactors in real time. SPR-based biosensors can be used in determining activity concentration, screening, and characterization in terms of both affinity and kinetics. The presence or level of one or more reference or non-reference biological activities can be measured according to cell-based assays, including assays that utilize a cellular binding partner of a selected anti-IL-18BP antibody functionally coupled to a readout or indicator, such as a fluorescent or luminescent indicator of biological activity, as described herein.

[0245] In certain embodiments, as described above, the composition is substantially endotoxin-free, e.g., about or at least about 95% endotoxin-free, about or at least about 99% endotoxin-free, or about or at least about 99.99% endotoxin-free. The presence of endotoxin can be detected according to routine techniques in the art, as described herein. In certain embodiments, the composition is produced from eukaryotic cells, such as mammalian or human cells, in a substantially serum-free medium. In certain embodiments, as described herein, the composition has an endotoxin content of less than about 10 EU / mg antibody, or less than about 5 EU / mg antibody, or less than about 3 EU / mg antibody, or less than about 1 EU / mg antibody.

[0246] In certain embodiments, the composition comprises less than about 10% w / w high molecular weight aggregates, or less than about 5% w / w high molecular weight aggregates, or less than about 2% w / w high molecular weight aggregates, or less than about 1% w / w high molecular weight aggregates.

[0247] Also included are protein-based analytical assays and methods that can be used to assess, among other characteristics, protein purity, size, solubility, and degree of aggregation. Protein purity can be assessed in a number of ways. For example, purity can be assessed based on primary structure, higher-order structure, size, charge, hydrophobicity, and glycosylation. Examples of methods for assessing primary structure include N- and C-terminal sequencing and peptide mapping (see, e.g., Allen et al., Biologicals. 24:255-275, 1996). Exemplary methods for assessing conformation include circular dichroism (see, e.g., Kelly et al., Biochim Biophys Acta. 1751:119-139, 2005), fluorescence spectroscopy (see, e.g., Meagher et al., J. Biol. Chem. 273:23283-89, 1998), FT-IR, amide hydrogen-deuterium exchange kinetics, differential scanning calorimetry, NMR spectroscopy, and immunoreactivity with conformation-sensitive antibodies. Conformation can also be assessed as a function of various parameters, such as pH, temperature, or added salt. Exemplary methods for assessing protein characteristics, such as size, include analytical ultracentrifugation and size-exclusion HPLC (SEC-HPLC), and exemplary methods for measuring charge include ion-exchange chromatography and isoelectric focusing. Hydrophobicity can be assessed, for example, by reverse-phase HPLC and hydrophobic interaction chromatography HPLC. Glycosylation can affect pharmacokinetics (e.g., clearance), conformation or stability, receptor binding, and protein function and can be assessed, for example, by mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy.

[0248] As noted above, certain embodiments include the use of SEC-HPLC to evaluate protein characteristics, such as purity, size (e.g., size homogeneity), or degree of aggregation, and / or to purify proteins, among other uses. SEC also includes gel filtration chromatography (GFC) and gel permeation chromatography (GPC) and refers to a chromatographic method in which molecules in solution are separated in a porous material based on their size, or more specifically, their hydrodynamic volume, diffusion coefficient, and / or surface properties. This process is generally used to separate biological molecules and determine the molecular weight and molecular weight distribution of polymers. Typically, a biological or protein sample (e.g., a protein extract produced according to the protein expression methods provided herein and known in the art) is loaded into a selected size-exclusion column with a defined stationary phase (porous material), preferably a phase that does not interact with the proteins in the sample. In certain aspects, the stationary phase is composed of inert particles packed in a dense three-dimensional matrix within a glass or steel column. The mobile phase can be pure water, an aqueous buffer, an organic solvent, or a mixture thereof. Stationary phase particles typically have small pores and / or channels that allow only molecules below a certain size to enter. Larger particles are therefore excluded from these pores and channels, and their limited interaction with the stationary phase leads them to elute as a "fully excluded" peak at the beginning of the experiment. Smaller molecules can fit into the pores and are removed from the flowing mobile phase; the time they spend immobilized in the stationary phase pores depends, in part, on how far into the pores they penetrate. Their removal from the mobile phase flow causes them to elute for longer periods from the column, resulting in separation between particles based on differences in their size. A given size-exclusion column has a range of molecular weights that can be separated. Overall, molecules larger than the upper limit are not captured by the stationary phase, while molecules smaller than the lower limit enter the solid phase completely and elute as a single band, with molecules within the range eluting at different rates defined by their properties, such as hydrodynamic volume.For examples of these methods in practice with pharmaceutical proteins, see Bruner et al., Journal of Pharmaceutical and Biomedical Analysis. 15:1929-1935, 1997.

[0249] Protein purity for clinical use has also been discussed, for example, by Anicetti et al. (Trends in Biotechnology. 7:342-349, 1989). More recent technologies for analyzing protein purity include, but are not limited to, the LabChip GXII, an automated platform for rapid analysis of proteins and nucleic acids, which provides high-throughput analysis of protein titer, size, and purity analysis. In certain non-limiting embodiments, clinical-grade proteins, such as protein fragments and antibodies, can be obtained by utilizing a combination of chromatographic materials in at least two orthogonal steps, among other methods (see, e.g., Therapeutic Proteins: Methods and Protocols. Vol. 308, Eds., Smales and James, Humana Press Inc., 2005). Typically, protein drugs (e.g., antibodies and antigen-binding fragments) are substantially endotoxin-free, as measured according to techniques known in the art and described herein.

[0250] Protein solubility assays are also included. Such assays can be used, for example, to determine optimal growth and purification conditions for recombinant production, optimize buffer selection, and optimize the selection of antibodies or antigen-binding fragments thereof. Solubility or aggregation can be assessed according to various parameters, including temperature, pH, salt, and the presence or absence of other additives. Examples of solubility screening assays include, but are not limited to, microplate-based methods that measure protein solubility using turbidity or other measures as an endpoint, high-throughput assays for analyzing the solubility of purified recombinant proteins (see, e.g., Stenvall et al., Biochim Biophys Acta. 1752:6-10, 2005), assays that use structural complementation of genetic marker proteins to monitor and measure protein folding and solubility in vivo (see, e.g., Wigley et al., Nature Biotechnology. 19:131-136, 2001), and electrochemical screening of recombinant protein solubility in E. coli using scanning electrochemical microscopy (SECM) (see, e.g., Nagamine et al., Biotechnology and Bioengineering. 96:1008-1013, 2006), among others. Antibodies with increased solubility (or reduced aggregation) can be identified or selected according to routine techniques in the art, including simple in vivo assays for protein solubility (see, e.g., Maxwell et al., Protein Sci. 8:1908-11, 1999).

[0251] Protein solubility and aggregation can also be measured by dynamic light scattering techniques. Aggregation is a general term that encompasses several types of interactions or characteristics, including soluble / insoluble, covalent / non-covalent, reversible / irreversible, and native / denatured interactions and characteristics. For protein therapeutics, the presence of aggregates is typically considered undesirable due to concerns that aggregates may cause immunogenic reactions (e.g., small aggregates) or adverse events upon administration (e.g., particulates). Dynamic light scattering refers to a technique that can be used to determine the size distribution profile of small particles in suspension or polymers in solution, such as proteins. This technique, also referred to as photon correlation spectroscopy (PCS) or quasi-elastic light scattering (QELS), uses scattered light to measure the rate of diffusion of protein particles. Fluctuations in scattering intensity can be observed due to Brownian motion of molecules and particles in solution. This motion data can be conventionally processed to derive a size distribution for the sample, in which the size is given by the Stokes radius or hydrodynamic radius of the protein particles. Hydrodynamic size depends on both mass and shape (composition). Dynamic light scattering can detect the presence of very small amounts of aggregated protein (<0.01% by weight) even in samples containing a wide range of masses. It can also be used, for example, to compare the stability of different formulations, including applications that rely on real-time monitoring of changes at elevated temperatures. Thus, certain embodiments include the use of dynamic light scattering to analyze the solubility and / or presence of aggregates in samples containing antibodies of the present disclosure.

[0252] Although the foregoing embodiments have been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in view of the teachings of the present disclosure that certain changes and modifications can be made without departing from the spirit or scope of the appended claims. The following examples are provided by way of illustration only, and not by way of limitation. Those of ordinary skill in the art will readily recognize a variety of non-critical parameters that can be changed or modified to yield essentially similar results.

[0253] Enumerated Embodiments The following non-limiting enumerated embodiments are provided by way of example.

[0254] Embodiment I-1. 1. An isolated antibody, or antigen-binding fragment thereof, that binds to interleukin-18 binding protein (IL-18BP), wherein the at least one antibody or antigen-binding fragment thereof is: V of a complementarity-determining region selected from Table A1 H CDR1 sequence, V H CDR2 sequence, and V H The heavy chain variable region (V H ), and variants thereof that specifically bind to IL-18BP; and V of a complementarity-determining region selected from Table A1 L CDR1 sequence, V L CDR2 sequence, and V L The light chain variable region (V L ), and variants thereof, which specifically bind to IL-18BP, or antigen-binding fragments thereof.

[0255] Embodiment I-2. The VHCDR1, VHCDR2, and VHCDR3 sequences are TFX1X2X3X4X5H and IX6X7X8X9X, respectively. 10 X 11 X 12 X 13 X 14 X 15 AQKFQG, and X 16 X 17 X 18 X 19 X 20 X 21 X 22 DY, and the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each contain X 23 X 24 X 25 X 26 X 27 X 28 X 29 X30 WX 31 A, X 32 X 33 X 34 X 35 X 36 X 37 X 38 , and QX 39 X 40 X 41 SFPYX 42 (see Table E11 for definition of "X" residues), V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 4 to 6, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 28 to 30, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 31 to 33, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 34 to 36, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 34 to 39, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 40 to 42, respectively; V H CDR1 sequence, VH CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 46 to 48, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 52 to 54, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 58 to 60, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 64 to 66, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 70 to 72, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 109 to 111, respectively, and VL CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 124 to 126, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 130 to 132, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 136 to 138, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 139 to 141, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 142 to 144, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 148 to 150, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 154 to 156, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 160 to 162, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 172 to 174, respectively; V HCDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 190 to 192, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 193 to 195, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 199 to 201, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; V H CDR1 sequence, V H CDR2 sequence, and V HThe CDR3 sequences comprise SEQ ID NOs: 205 to 207, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 208-210, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 211 to 213, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 214-216, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 217 to 219, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 223 to 225, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 229 to 231, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 232 to 234, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 235 to 237, respectively, and V L CDR1 sequence, VL CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 241 to 243, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 244 to 246, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 247 to 249, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 250 to 252, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 253 to 255, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 256 to 258, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 265 to 267, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 268-270, respectively; or V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 271 to 273, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V LThe isolated antibody, or antigen-binding fragment thereof, of embodiment I-1, wherein the CDR3 sequences comprise SEQ ID NOs: 274-276, respectively.

[0256] Embodiment I-3. V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V H The isolated antibody, or antigen-binding fragment thereof, of embodiment I-1 or I-2, having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

[0257] Embodiment I-4. V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V L The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-3, having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

[0258] Embodiment I-5. V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 286; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 288; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 292; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 294; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 296; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 298; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 352; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 353, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 354; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 355, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 356; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 357, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 358; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 359, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 360; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 361, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 362; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 368; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 369, and V L The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-4, comprising a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 370.

[0259] Embodiment I-6. An isolated antibody, or antigen-binding fragment thereof, according to any one of embodiments I-1 to I-5, wherein the antibody binds to an epitope comprising I97 and V153 according to UniProt:O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO:1, or I67 and V123 as set forth in SEQ ID NO:2).

[0260] Embodiment I-7. An isolated antibody or antigen-binding fragment thereof, wherein the antibody binds to interleukin-18 binding protein (IL-18BP) at an epitope comprising I97 and V153 according to UniProt:O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO:1, or I67 and V123 as set forth in SEQ ID NO:2).

[0261] Embodiment I-8. The VHCDR1, VHCDR2, and VHCDR3 sequences are TFX1X2X3X4X5H and IX6X7X8X9X, respectively. 10 X 11 X 12 X 13 X 14 X 15 AQKFQG, and X 16 X 17 X18 X 19 X 20 X 21 X 22 DY, and the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each contain X 23 X 24 X 25 X 26 X 27 X 28 X 29 X 30 WX 31 A, X 32 X 33 X 34 X 35 X 36 X 37 X 38 , and QX 39 X 40 X 41 SFPYX 42 (see Table E11 for definition of "X" residues), V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 4-6, respectively; or V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 265 to 267, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L The isolated antibody, or antigen-binding fragment thereof, of embodiment I-7, wherein the CDR3 sequences comprise SEQ ID NOs: 268-270, respectively.

[0262] Embodiment I-9. V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and V L The isolated antibody of embodiment I-8, or an antigen-binding fragment thereof, comprising a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 368.

[0263] Embodiment I-10. An isolated antibody, or antigen-binding fragment thereof, wherein the antibody binds to interleukin-18 binding protein (IL-18BP) and the antibody interferes with the binding of IL-18 to IL-18BP.

[0264] Embodiment I-11. The isolated antibody, or antigen-binding fragment thereof, of embodiment I-10, wherein the antibody binds to a conformational epitope of IL-18BP.

[0265] Embodiment I-12. The isolated antibody, or antigen-binding fragment thereof, of embodiment I-10 or I-11, wherein the conformational epitope of IL-18BP comprises two or more amino acid residues selected from the group consisting of K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO: 372.

[0266] Embodiment I-13. The isolated antibody, or antigen-binding fragment thereof, according to any one of embodiments I-10 to I-12, wherein the conformational epitope of IL-18BP comprises amino acid residues K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO: 372.

[0267] Embodiment I-14. The isolated antibody, or antigen-binding fragment thereof, of embodiment I-10, wherein the antibody binds to a linear epitope of IL-18BP.

[0268] Embodiment I-15. The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-10 to I-14, wherein the antibody binds to the binding interface between IL-18 and mature IL-18BP.

[0269] Embodiment I-16. The isolated antibody of embodiment I-15, or an antigen-binding fragment thereof, wherein the antibody binds to amino acid residues R61, Y69, and R131 of SEQ ID NO: 372.

[0270] Embodiment I-17. V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and V L The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-12 to I-16, comprising a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334.

[0271] Embodiment I-18. An isolated antibody, or antigen-binding fragment thereof, according to any one of embodiments I-1 to I-17, wherein the antibody binds to human IL-18BP and cynomolgus IL-18BP, but does not bind (specifically or substantially) to mouse IL-18BP.

[0272] Embodiment I-19. The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-17, wherein the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

[0273] Embodiment I-20. The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-19, wherein the antibody binds to the binding interface between IL-18 and mature IL-18BP.

[0274] Embodiment I-21. The binding affinity between IL-18 and IL-18BP (K D 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM or less, and optionally with a binding affinity of about 1 pm to about 650 pm, or about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM or less.

[0275] Embodiment I-22. The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-21, which is an IL-18BP antagonist, antagonizing the binding activity between IL-18BP and IL-18.

[0276] Embodiment I-23. The isolated antibody of embodiment I-22, or an antigen-binding fragment thereof, wherein the antibody blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signaling, including induction of IFN-gamma, CXCL10 and / or TNFα.

[0277] Embodiment I-24. An isolated antibody, or antigen-binding fragment thereof, according to any one of embodiments I-1 to I-8, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or hybrids and / or variants thereof.

[0278] Embodiment I-25. The isolated antibody or antigen-binding fragment thereof of embodiment I-24, comprising an IgG Fc domain with enhanced effector function in humans, optionally an IgG1 or IgG3 Fc domain.

[0279] Embodiment I-26. The isolated antibody or antigen-binding fragment thereof of embodiment I-24, wherein the antibody comprises an IgG Fc domain with reduced effector function in humans, optionally an IgG2 or IgG4 Fc domain.

[0280] Embodiment I-27. The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-26, which is a monoclonal antibody.

[0281] Embodiment I-28. The isolated antibody, or antigen-binding fragment thereof, of any one of embodiments I-1 to I-27, which is a humanized antibody.

[0282] Embodiment I-29. The isolated antibody or antigen-binding fragment thereof of any one of embodiments I-1 to I-28, wherein the antibody is selected from an Fv fragment, a single-chain Fv (scFv) polypeptide, an adnectin, an anticalin, an aptamer, an avimer, a camelid antibody, a designed ankyrin repeat protein (DARPin), a minibody, a nanobody, and a unibody.

[0283] Embodiment I-30. An isolated polynucleotide encoding the isolated anti-IL-18BP antibody or antigen-binding fragment thereof according to any one of embodiments I-1 to I-29, an expression vector comprising the isolated polynucleotide, or an isolated host cell comprising the vector.

[0284] Embodiment I-31. A pharmaceutical composition comprising the isolated anti-IL-18BP antibody or antigen-binding fragment thereof according to any one of embodiments I-1 to I-29, and a pharmaceutically acceptable carrier.

[0285] Embodiment I-32. The pharmaceutical composition of embodiment I-31, wherein the composition has a purity of at least about 80%, 85%, 90%, 95%, 98%, or 99% on a protein basis with respect to at least one antibody or antigen-binding fragment, and is substantially free of aggregates and endotoxin.

[0286] Embodiment I-33. The pharmaceutical composition of embodiment I-31 or I-32, wherein the composition is optionally a sterile injectable solution suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

[0287] Embodiment I-34. A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of embodiments I-31 to I-33.

[0288] Embodiment I-35. The method of embodiment I-34, wherein the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, optionally a proliferative disease or disorder selected from a lymphoproliferative disorder, a myeloproliferative disorder, a proliferative enterocolitis, a proliferative diabetic retinopathy, and a proliferative kidney disease.

[0289] Embodiment I-36. The method of embodiment I-35, wherein the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18.

[0290] Embodiment I-37. Cancers include bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (liver cell carcinoma), sarcoma, B-cell malignancies, breast cancer, The method of embodiment I-35 or I-36, wherein the cancer is selected from one or more of: ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, urothelial cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer.

[0291] Embodiment I-38. The method according to any one of embodiments I-34 to I-37, comprising administering a pharmaceutical composition (comprising an anti-IL18BP antibody or an antigen-binding fragment thereof) in combination with IL-18.

[0292] Embodiment I-39. The method of any one of embodiments I-35 to I-38, comprising administering a pharmaceutical composition (comprising an anti-IL18BP antibody or antigen-binding fragment thereof) in combination with an immune checkpoint modulator selected from an antagonist of an inhibitory immune checkpoint molecule, and an agonist of a stimulatory immune checkpoint molecule.

[0293] Embodiment I-40. The method of embodiment I-39, wherein the immune checkpoint modulator is a polypeptide, optionally an antibody or antigen-binding fragment thereof, or a ligand, or a small molecule.

[0294] Embodiment I-41. The method of embodiment I-39 or I-40, wherein the inhibitory immune checkpoint molecule is selected from one or more of Programmed Death-Ligand 1 (PD-L1), Programmed Death 1 (PD-1), Programmed Death-Ligand 2 (PD-L2), Cytotoxic T-Lymphocyte-Associated protein 4 (CTLA-4), Indoleamine 2,3-dioxygenase (IDO), Tryptophan 2,3-dioxygenase (TDO), T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3), Lymphocyte Activation Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T Lymphocyte Attenuator (BTLA), CD160, Herpes Virus Entry Mediator (HVEM), and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0295] Embodiment I-42. the antagonist is an antibody or antigen-binding fragment or small molecule that specifically binds to PD-L1 and / or PD-L2, an antagonist of PD-L1 and / or PD-L2 optionally chosen from one or more of atezolizumab (MPDL3280A), avelumab (MSB0010718C) and durvalumab (MEDI4736), optionally in which the cancer is selected from one or more of colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer and renal cell carcinoma; the antagonist is a PD-1 antagonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally in which the PD-1 antagonist is nivolumab, and the cancer is optionally chosen from one or more of Hodgkin lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer; the PD-1 antagonist is pembrolizumab, and the cancer is optionally selected from one or more of melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial cancer; the antagonist is a CTLA-4 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, optionally in which the cancer is selected from melanoma, prostate cancer, lung cancer, and bladder cancer; the antagonist is an IDO antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-l-tryptophan (1MT), β-carboline (norharman; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, wherein the cancer is optionally selected from one or more of metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or malignant brain tumor; the antagonist is a TDO antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to TDO, 680C91 and LM10; The antagonist is a TIM-3 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to TIM-3; the antagonist is a LAG-3 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to LAG-3, and BMS-986016; The antagonist is a VISTA antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to VISTA; the antagonist is an antagonist of BTLA, CD160 and / or HVEM optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to BTLA, CD160 and / or HVEM; The method of embodiment I-41, wherein the antagonist is a TIGIT antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to TIGIT.

[0296] Embodiment I-43. The method of embodiment I-39 or I-40, wherein the stimulatory immune checkpoint molecule is selected from one or more of OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

[0297] Embodiment I-44. the agonist is an OX40 agonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L,...

Claims

1. 1. An isolated antibody, or antigen-binding fragment thereof, that binds to interleukin-18 binding protein (IL-18BP), wherein said at least one antibody or antigen-binding fragment thereof comprises: V of a complementarity determining region selected from Table A1 Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The heavy chain variable region (V) containing the CDR3 sequence Ｈ ), and variants thereof that specifically bind to IL-18BP; and V of a complementarity determining region selected from Table A1 Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ The light chain variable region (V) containing the CDR3 sequence Ｌ ), and variants thereof that specifically bind to IL-18BP, or antigen-binding fragments thereof.

2. The VHCDR1 sequence, VHCDR2 sequence, and VHCDR3 sequence are each １ X ２ X ３ X ４ X ５ H, IX ６ X ７ X ８ X ９ X １０ X １１ X １２ X １３ X １４ X １５ AQKFQG, and X １６ X １７ X １８ X １９ X ２０ X ２１ X ２２ DY, wherein the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each comprise X ２３ X ２４ X ２５ X ２６ X ２７ X ２８ X ２９ X ３０ WX ３１ A, X ３２ X ３３ X ３４ X ３５ X ３６ X ３７ X ３８ , and QX ３９ X ４０ X ４１ SFPYX ４２ (see Table E11 for definition of "X" residues), The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 1-3, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 4-6, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 25-27, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 28-30, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 31-33, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 34-36, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 34-39, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 40-42, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 43-45, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 46-48, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 49-51, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 52-54, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 55-57, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 58-60, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 61-63, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 64-66, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 67-69, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 70-72, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 109-111, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 115-117, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 121-123, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 124-126, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 127-129, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 130-132, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 133-135, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 136-138, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 139-141, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 142-144, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 145-147, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 148-150, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 151-153, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 154-156, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 157-159, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 160-162, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 163-165, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 172-174, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 175-177, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 181-183, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 184-186, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 187-189, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 190-192, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 193-195, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 199-201, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 205-207, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs:208-210, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 211-213, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs:214-216, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 217-219, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 223-225, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 229-231, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 232-234, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 235-237, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 241-243, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 244-246, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 250-252, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 253-255, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 256-258, respectively; The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 265-267, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 268-270, respectively; or The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 271-273, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, wherein the CDR3 sequences comprise SEQ ID NOs: 274-276, respectively.

3. The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, said V Ｈ 3. The isolated antibody, or antigen-binding fragment thereof, of claim 1 or 2, wherein said antibody has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

4. The V Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, said V Ｌ 4. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 3, wherein:

5. The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:278; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:286; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:288; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:292; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:294; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:296; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:298; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 352; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 353, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 354; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 355, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 356; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 357, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 358; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 359, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 360; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 361, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 362; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 368; or The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 369, and Ｌ The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 4, comprising a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:

370.

6. 6. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 5, wherein the antibody binds to an epitope comprising I97 and V153 according to UniProt: O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO: 1, or I67 and V123 as set forth in SEQ ID NO: 2).

7. An isolated antibody or antigen-binding fragment thereof, wherein the antibody binds to interleukin-18 binding protein (IL-18BP) at an epitope comprising I97 and V153 according to UniProt: O95998 numbering (alternatively, I95 and V151 as set forth in SEQ ID NO: 1, or I67 and V123 as set forth in SEQ ID NO: 2).

8. The VHCDR1 sequence, VHCDR2 sequence, and VHCDR3 sequence are each １ X ２ X ３ X ４ X ５ H, IX ６ X ７ X ８ X ９ X １０ X １１ X １２ X １３ X １４ X １５ AQKFQG, and X １６ X １７ X １８ X １９ X ２０ X ２１ X ２２ DY, wherein the VLCDR1 sequence, the VLCDR2 sequence, and the VLCDR3 sequence each comprise X ２３ X ２４ X ２５ X ２６ X ２７ X ２８ X ２９ X ３０ WX ３１ A, X ３２ X ３３ X ３４ X ３５ X ３６ X ３７ X ３８ , and QX ３９ X ４０ X ４１ SFPYX ４２ (see Table E11 for definition of "X" residues), The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 1-3, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 4-6, respectively; or The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 265-267, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ 8. The isolated antibody, or antigen-binding fragment thereof, of claim 7, wherein the CDR3 sequences comprise SEQ ID NOs: 268-270, respectively.

9. The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:278; The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and Ｌ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; or The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 367, and Ｌ 9. The isolated antibody, or antigen-binding fragment thereof, of claim 8, comprising a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:

368.

10. An isolated antibody, or antigen-binding fragment thereof, which binds to interleukin-18 binding protein (IL-18BP), and which interferes with the binding of IL-18 to IL-18BP.

11. The isolated antibody, or antigen-binding fragment thereof, of claim 10, wherein the antibody binds to a conformational epitope of IL-18BP.

12. 12. The isolated antibody, or antigen-binding fragment thereof, according to claim 10 or 11, wherein the conformational epitope of IL-18BP comprises two or more amino acid residues selected from the group consisting of K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO:

372.

13. 13. The isolated antibody, or antigen-binding fragment thereof, according to any one of claims 10 to 12, wherein the conformational epitope of IL-18BP comprises amino acid residues K32, T40, S60, R61, S66, Y69, R91, R93, T96, K102, R131, and H132 of SEQ ID NO:

372.

14. 11. The isolated antibody, or antigen-binding fragment thereof, of claim 10, wherein the antibody binds to a linear epitope of IL-18BP.

15. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 10 to 14, wherein the antibody binds to the binding interface between IL-18 and mature IL-18BP.

16. 16. The isolated antibody, or antigen-binding fragment thereof, of claim 15, wherein the antibody binds to amino acid residues R61, Y69, and R131 of SEQ ID NO:

372.

17. The V Ｈ comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and Ｌ 17. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 12 to 16, comprising a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:

334.

18. 18. The isolated antibody, or antigen-binding fragment thereof, according to any one of claims 1 to 17, wherein the antibody binds to human IL-18BP and cynomolgus monkey IL-18BP, but does not bind (specifically or substantially) to mouse IL-18BP.

19. The isolated antibody, or antigen-binding fragment thereof, according to any one of claims 1 to 17, wherein the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

20. 20. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 19, wherein the antibody binds to the binding interface between IL-18 and mature IL-18BP.

21. The binding affinity between IL-18 and IL-18BP (K Ｄ 21. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 20, which binds to IL-18BP with a binding affinity of greater than about 650 pM, optionally between about 1 pm and about 650 pm, or about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM or less.

22. 22. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 21, which is an IL-18BP antagonist that antagonizes the binding activity between IL-18BP and IL-18.

23. 23. The isolated antibody, or antigen-binding fragment thereof, of claim 22, wherein the antibody blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signaling, including induction of IFN-gamma, CXCL10 and / or TNFα.

24. 9. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 8, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or hybrids and / or variants thereof.

25. 25. The isolated antibody, or antigen-binding fragment thereof, of claim 24, comprising an IgG Fc domain, optionally an IgG1 or IgG3 Fc domain, that has enhanced effector function in humans.

26. 25. The isolated antibody, or antigen-binding fragment thereof, of claim 24, comprising an IgG Fc domain, optionally an IgG2 or IgG4 Fc domain, that has reduced effector function in humans.

27. 27. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 26, which is a monoclonal antibody.

28. 28. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 27, which is a humanized antibody.

29. 29. The isolated antibody, or antigen-binding fragment thereof, of any one of claims 1 to 28, selected from an Fv fragment, a single-chain Fv (scFv) polypeptide, an adnectin, anticalin, an aptamer, an avimer, a camelid antibody, a designed ankyrin repeat protein (DARPin), a minibody, a nanobody, and a unibody.

30. 30. An isolated polynucleotide encoding the isolated anti-IL-18BP antibody or antigen-binding fragment thereof according to any one of claims 1 to 29, an expression vector comprising said isolated polynucleotide, or an isolated host cell comprising said vector.

31. A pharmaceutical composition comprising the isolated anti-IL-18BP antibody or antigen-binding fragment thereof according to any one of claims 1 to 29 and a pharmaceutically acceptable carrier.

32. 32. The pharmaceutical composition of claim 31, wherein the composition has a purity of at least about 80%, 85%, 90%, 95%, 98%, or 99% on a protein basis with respect to the at least one antibody or antigen-binding fragment, and is substantially free of aggregates and endotoxins.

33. 33. The pharmaceutical composition of claim 31 or 32, wherein the composition is a sterile injectable solution, optionally suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

34. 34. A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of claims 31 to 33.

35. 35. The method of claim 34, wherein the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, optionally a proliferative disease or disorder selected from lymphoproliferative disorders, myeloproliferative disorders, proliferative enterocolitis, proliferative diabetic retinopathy, and proliferative kidney disease.

36. 36. The method of claim 35, wherein the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18.

37. The cancers include bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (liver cell carcinoma), sarcoma, B-cell malignancies, 37. The method of claim 35 or 36, wherein the tumor is selected from one or more of: breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, urothelial cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer.

38. The method of any one of claims 34 to 37, comprising administering the pharmaceutical composition (comprising the anti-IL18BP antibody or antigen-binding fragment thereof) in combination with IL-18.

39. 39. The method of any one of claims 35 to 38, comprising administering the pharmaceutical composition (comprising the anti-IL18BP antibody or antigen-binding fragment thereof) in combination with an immune checkpoint modulator selected from an antagonist of an inhibitory immune checkpoint molecule, and an agonist of a stimulatory immune checkpoint molecule.

40. 40. The method of claim 39, wherein the immune checkpoint modulator is a polypeptide, optionally an antibody or antigen-binding fragment thereof, or a ligand, or a small molecule.

41. The inhibitory immune checkpoint molecules include Programmed Death-Ligand 1 (PD-L1), Programmed Death 1 (PD-1), Programmed Death-Ligand 2 (PD-L2), Cytotoxic T-Lymphocyte-Associated protein 4 (CTLA-4), Indoleamine 2,3-dioxygenase (IDO), Tryptophan 2,3-dioxygenase (TDO), T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3), Lymphocyte Activation Regulatory Factor (LIF-FRF) and Lymphocyte Activation Factor (LIF-FRF). The method of claim 39 or 40, wherein the target gene is selected from one or more of Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T lymphocyte attenuator (BTLA), CD160, Herpes Virus Entry Mediator (HVEM), and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

42. the antagonist is an antagonist of PD-L1 and / or PD-L2 optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to PD-L1 and / or PD-L2, atezolizumab (MPDL3280A), avelumab (MSB0010718C), and durvalumab (MEDI4736), and optionally the cancer is selected from one or more of colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer, and renal cell carcinoma; the antagonist is a PD-1 antagonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally the PD-1 antagonist is nivolumab, and the cancer is optionally chosen from one or more of Hodgkin's lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer; the PD-1 antagonist is pembrolizumab, and the cancer is optionally selected from one or more of melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial cancer; the antagonist is a CTLA-4 antagonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, and optionally the cancer is chosen from melanoma, prostate cancer, lung cancer, and bladder cancer; the antagonist is an IDO antagonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-1-tryptophan (1MT), β-carboline (norharman; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, and the cancer is optionally chosen from one or more of metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or malignant brain tumor; the antagonist is a TDO antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to TDO, 680C91, and LM10; the antagonist is a TIM-3 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to TIM-3; the antagonist is a LAG-3 antagonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule that specifically binds to LAG-3, and BMS-986016; the antagonist is a VISTA antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to VISTA; the antagonist is an antagonist of BTLA, CD160 and / or HVEM, optionally chosen from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to BTLA, CD160 and / or HVEM; 42. The method of claim 41, wherein the antagonist is a TIGIT antagonist optionally selected from one or more of an antibody or antigen-binding fragment or a small molecule that specifically binds to TIGIT.

43. 41. The method of claim 39 or 40, wherein the stimulatory immune checkpoint molecule is selected from one or more of OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

44. the agonist is an OX40 agonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L, and GSK3174998; the agonist is a CD40 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD40, CP-870,893, dacetuzumab, Chi Lab 7 / 4, ADC-1013, and rhCD40L, and the cancer is optionally selected from one or more of melanoma, pancreatic cancer, mesothelioma, and hematological cancers, optionally lymphomas, e.g., non-Hodgkin's lymphoma; the agonist is a GITR agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to GITR, INCAGN01876, DTA-1, and MEDI1873; the agonist is a CD137 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD137, utomilumab, and 4-1BB ligand; the agonist is a CD27 agonist optionally chosen from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD27, varlilumab, and CDX-1127 (1F5); the agonist is a CD28 agonist optionally selected from one or more of an antibody or antigen-binding fragment or small molecule or ligand that specifically binds to CD28, and TAB08; and / or 44. The method of claim 43, wherein the agonist is an HVEM agonist optionally selected from one or more of an antibody or antigen-binding fragment, or a small molecule, or a ligand that specifically binds to HVEM.

45. The method of any one of claims 35 to 44, comprising administering said pharmaceutical composition (comprising said anti-IL18BP antibody or antigen-binding fragment thereof) in combination with at least one chemotherapeutic agent.

46. 46. ​​The method of claim 45, wherein the at least one chemotherapeutic agent is selected from one or more of an alkylating agent, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor (type 1 or type II), and an anti-microtubule agent.

47. said alkylating agents are selected from one or more of nitrogen mustards (optionally mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosoureas (optionally N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, and streptozotocin), tetrazines (optionally dacarbazine, mitozolomide, and temozolomide), aziridines (optionally thiotepa, mitomycin, and diaziquone (AZQ)), cisplatin and its derivatives (optionally carboplatin and oxaplatin), and non-classical alkylating agents (optionally procarbazine and hexamethylmelamine); the antimetabolite is selected from one or more of antifolates (optionally methotrexate and pemetrexed), fluoropyrimidines (optionally 5-fluorouracil and capecitabine), deoxynucleoside analogs (optionally ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (optionally thioguanine and mercaptopurine); said cytotoxic antibiotic is selected from one or more of anthracyclines (optionally doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, and mitoxantrone), bleomycin, mitomycin C, mitoxantrone, and actinomycin; the topoisomerase inhibitor is selected from one or more of camptothecin, irinotecan, topotecan, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, mervalone, and aclarubicin; and / or 47. The method of claim 46, wherein the anti-microtubule agent is selected from one or more of taxanes (optionally paclitaxel and docetaxel) and vinca alkaloids (optionally vinblastine, vincristine, vindesine, vinorelbine).

48. 35. The method of claim 34, wherein the disease or condition is myelodysplastic syndrome (MDS).

49. 35. The method of claim 34, wherein the disease or condition is an infectious disease.

50. 50. The method of claim 49, wherein the infectious disease is selected from viral, bacterial, fungal (optionally yeast), and protozoal infections.

51. The method of any one of claims 48 to 50, comprising administering said pharmaceutical composition (comprising said anti-IL18BP antibody or antigen-binding fragment thereof) in combination with IL-18.

52. 1. A method of screening an anti-IL-18BP antibody or antigen-binding fragment thereof for the ability to block or inhibit binding between IL-18 and IL-18BP, comprising: a. i. IL-18BP alone, and ii. determining the binding affinity of the antibody or antigen-binding fragment thereof to an IL-18-reduced fusion protein, the IL-18-reduced fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and an optional protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein is bound to the IL-18BP portion of the fusion protein so as to sterically block the IL-18 binding site of the IL-18BP portion of the fusion protein; b. comparing the binding affinity of (i) with the binding affinity of (ii); and c) identifying or selecting said antibody or antigen-binding fragment thereof as being capable of blocking or inhibiting the binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii).

53. The method of claim 52, wherein the IL-18 and IL-18BP are mouse IL-18 and IL-18BP.

54. The method of claim 52, wherein the IL-18 and IL-18BP are human IL-18 and IL-18BP.

55. 55. The method of any one of claims 52 to 54, wherein the IL-18-reduced fusion protein comprises, from N-terminal to C-terminal direction, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP.

56. 56. The method of claim 55, wherein the IL-18-low fusion protein comprises an amino acid sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

57. A reduced IL-18 fusion protein comprising, in the N-terminal to C-terminal direction, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP.

58. 58. The IL-18-reduced fusion protein of claim 57, comprising an amino acid sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

59. A method of stimulating an immune response in a subject in need thereof, comprising administering to said subject the pharmaceutical composition of any one of claims 31 to 33.

60. 60. The method of claim 59, wherein the immune response is an IL-18 mediated immune response.

61. 61. The method of claim 60, wherein the IL-18 mediated immune response comprises the induction of IFN-gamma, CXCL10, and / or TNFα in the subject in need thereof.

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