Antiphosphocholine antibody and method of use thereof

Abstract

This disclosure provides polypeptides that specifically bind to phosphocholine (PC). Also provided are pharmaceutical compositions comprising these polypeptides, nucleic acids encoding these polypeptides, expression vectors and host cells for producing these polypeptides, and methods for treating a target using these polypeptides.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 452,798, filed on 17 March 2023, the entire disclosure of which is incorporated herein by reference.

[0002] Sequence listing reference This application includes a sequence listing filed electronically in ST.26 format, which is incorporated herein by reference in its entirety. The ST.26 copy was created on 15 March 2024, is named "208485_seqlist.xml", and is 644,762 bytes in size.

[0003] This disclosure relates to an antibody specific to phosphocholine (PC) and a method of using the same. [Background technology]

[0004] Phospholipids containing polyunsaturated fatty acids are highly susceptible to modification by reactive oxygen species (ROS) and non-radical oxidants. Oxidized lipids in cell membranes and circulation can have both beneficial and harmful effects on the human body. Endogenous oxidized phospholipids (OxPL) have been shown to function as stress signals, prompting phagocytic cells such as macrophages to respond by inducing inflammatory programs and eliminating the source of stress. However, excess oxidized phospholipid products are associated with chronic inflammation, as well as the pathogenesis of various cardiopulmonary disorders such as atherosclerosis and thrombosis, acute lung injury, and neurodegenerative processes.

[0005] Oxidized low-density lipoproteins (OxLDL), comprising several OxPL species, are found in atherosclerotic lesions and are detected at high concentrations in the serum of patients with hyperlipidemia, diabetes, and liver disease. The phosphocholine (PC) head group of OxPL has been shown to be important for phospholipid binding to intrinsic membrane proteins such as CD36 and the innate defense molecule C-reactive protein (CRP) (Non-patent Literature 1 and 2).

Prior Art Documents

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] Therefore, there is a need for therapies that include antibodies that bind to PC in order to target OxPL across a wide range of diseases.

Means for Solving the Problems

[0008] The present disclosure provides polypeptides (e.g., antibodies) that specifically bind to PC. Pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for producing these antibodies, and methods of treating a subject using these antibodies are also provided. The polypeptides provided herein are particularly advantageous because they bind to PC with high affinity, exhibit good stability, stress tolerance, and immunogenic profiles, and can be produced in high yields. The polypeptides provided herein are particularly useful for treating inflammatory disorders or degenerative diseases in a subject.

[0009] Accordingly, in one embodiment, the present disclosure provides an antibody that specifically binds to phosphocholine (PC), wherein the antibody comprises VH, which includes one of the VH amino acid sequences described in SEQ ID NOs. 1 to 180, consisting of CDRH1, CDRH2, and CDRH3 amino acid sequences, and VL, which includes one of the VL amino acid sequences described in SEQ ID NOs. 181 to 368, consisting of CDRL1, CDRL2, and CDRL3 amino acid sequences.

[0010] In one embodiment, the antibodies are, respectively, SEQ ID NOs: 1 and 181;2 and 182;3 and 183;4 and 184;5 and 185;1 and 184;6 and 184;1 and 186;7 and 187;8 and 188;9 and 189;10 and 190;4 and 191;11 and 192;12 and 193;13 and 194;14 and 195;15 and 196;16 and 184;17 and 197;18 and 198;1 and 199;19 and 200;20 and 201;21 and 202;22 and 203;23 and 20 4;24 and 205;25 and 184;4 and 206;26 and 207;27 and 208;28 and 209;9 and 210;29 and 211;30 and 212;31 and 213;32 and 214;33 and 215;34 and 216;17 and 217;1 and 218;35 and 184;36 and 184;35 and 219;35 and 220;37 and 184;38 and 221;39 and 222;9 and 223;9 and 224;40 and 184;41 and 225;8 and 226;42 and 227;35 and 228;43 and 229;4 and 230;9 and 231;8 and 232;1 and 233;8 and 234;44 and 184;1 and 235;45 and 236;30 and 237;46 and 184;47 and 238;48 and 239;1 and 230;49 and 190;44 and 240;50 and 184;51 and 241;52 and 242;53 and 243;54 and 184;55 and 244;56 and 245;57 and 246;58 and 247;59 and 248;60 and 249;8 and 250 ;61 and 184;35 and 251;62 and 252;1 and 253;63 and 254;64 and 181;65 and 255;66 and 256;67 and 257;68 and 198;35 and 258;69 and 259;70 and 260;71 and 261;72 and 262;73 and 263;74 and 264;75 and 265;76 and 266;77 and 267;4 and 205;78 and 268;1 and 269;27 and 269;79 and 269;80 and 270;81 and 269;22 and 269;82 and 269; 83 and 271; 84 and 272; 1 and 273; 8 and 274; 8 and 275; 22 and 276; 8 and 277; 85 and 278; 86 and 279; 87 and 269; 88 and 280; 89 and 269; 60 and 281; 90 and 269; 91 and 282; 22 and 283; 92 and 269; 93 and 284; 94 and 285; 95 and 286; 96 and 269; 73 and 287; 97 and 288; 68 and 289; 98 and 269; 99 and 290; 100 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304; 72 and 305; 114 and 306; 115 and 307; 116 and 269; 117 and 308;118 and 269;110 and 309;119 and 310;120 and 311;121 and 269;122 and 312;123 and 289;9 and 313;124 and 314;120 and 289;125 and 315;110 and 316;126 and 317;127 and 318;49 and 319;128 and 320;9 and 321;22 and 322;129 and 323;130 and 269;131 and 324;132 and 325;133 and 326;134 and 327;53 and 328 ;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340;149 and 341;150 and 342;151 and 343;152 and 344;94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 356; 166 and 357; 167 and 358; 110 and 334; 87 and 359; 1 The VH and VL amino acid sequences described in 68 and 360; 169 and 269; 170 and 269; 171 and 269; 172 and 361; 8 and 362; 173 and 304; 174 and 269; 1 and 363; 175 and 364; 176 and 365; 177 and 269; 178 and 366; 179 and 269; 180 and 367; or 89 and 368 include the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences.

[0011] In one embodiment, the antibodies are, respectively, SEQ ID NOs: 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381 , 428, and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 484; 385, 428, and 484; 372, 438, and 484; 388, 428, and 4 94;369, 439, and 484;389, 428, and 484;381, 428, and 484;376, 440, and 495;390, 441, and 484;370, 442, and 484;391, 428, and 484;392, 432, and 484;377, 443, and 484;393, 428, and 484;383, 444, and 489;394, 428, and 484;383, 444, and 496;376, 434, and 484;395, 445, and 497;396, 446, and 487;392, 42 8, and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484; 402, 436, and 484; 381, 451, and 484; 403, 428, and 484;369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428, and 497; 403, 453, and 484; 402, 454, and 484; 383, 434 , and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428, and 484; 415, 428, and 489; 369, 428, and 491; 403, 458 , and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512; 381, 428, and 493; 383, 470, and 501; 404, 471, and 484;418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515; 376, 428, and 504; 422, 469, and 4 84;369, 428, and 516;405, 460, and 484;413, 431, and 484;406, 429, and 484;383, 428, and 517;377, 437, and 518;369, 477, and 487;403, 464, and 484;369, 454, and 484;372, 478, and 519;423, 429, and 484;382, 434, and 484;424, 479, and and 484;376, 429, and 484;425, 428, and 484;403, 428, and 520;417, 460, and 484;409, 460, and 484;404, 452, and 484;383, 428, and 521;372, 480, and 493;369, 477, and 484;369, 481, and 484;384, 452, and 484;377, 482, and 484;410, 483, and 484; 384, 428, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or containing the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 400, 452, and 484.

[0012] In one embodiment, the antibodies are, respectively, SEQ ID NOs: 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523 , 601, and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 661; 523, 610, and 659; 523, 611, and 668; 523, 611, and 6 58; 523, 596, and 662; 523, 612, and 667; 523, 613, and 668; 523, 591, and 669; 523, 591, and 663; 523, 614, and 659; 532, 615, and 662; 523, 616, and 670; 523, 591, and 668; 523, 617, and 671; 533, 595, and 659; 523, 618, and 659; 534, 619, and 662; 535, 620, and 659; 536, 621, and 659; 537, 603, and 671; 523, 62 2, and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671; 539, 591, and 662; 523, 591, and 670; 523, 611, and 659;537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610, and 674; 523, 634, and 671; 545, 591, and 661; 523, 635 , and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616, and 660; 523, 602, and 677; 523, 616, and 659; 523, 630 , and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678; 571, 630, and 663; 572, 591, and 664; 573, 591, and 670;523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 523, 595, and 659; 523, 600, and 659; 523, 6 49, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 591, and 659; 584, 591, and 671; 523, 591, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; Containing the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 560, 591, and 659; 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or 523, 606, and 680.

[0013] In one embodiment, the antibodies are SEQ ID NOs: 369, 428, 484, 523, 591, and 656; 370, 429, 485, 523, 592, and 657; 371, 428, 486, 523, 593, and 658; 372, 428, 484, 523, 591, and 659; 373, 429, 484, 523, 594, and 659; 369, 428, 484, 523, 591, and 659; 374, 430, 484, 523, 591, and 659; 369, 428, 484, 523, 595, and 660; 375, 428, 487, 523 , 596, and 659; 376, 428, 484, 524, 591, and 659; 377, 428, 484, 523, 591, and 661; 378, 428, 484, 523, 591, and 662; 372, 428, 484, 523, 597, and 658; 379, 428, 484, 523, 598, and 662; 372, 428, 488, 525, 599, and 659; 376, 431, 484, 523, 600, and 663; 380, 432, 484, 523, 591, and 664; 372, 428, 489, 523, 601 , and 659;381,428,490,523,591, and 659;382,428,484,526,597, and 665;381,428,491,523,602, and 659;369,428,484,527,591, and 661;377,428,492,528,591, and 662;376,433,484,529,603, and 662;383,434,484,523,604, and 666;383,428,484,523,605, and 667;384,435,484,530,606, and 659;369, 436, 484, 523, 607, and 659;372, 437, 487, 523, 591, and 659;372, 428, 484, 523, 608, and 659;385, 428, 493, 523, 609, and 662;386, 428, 484, 531, 591, and 663;372, 437, 484, 523, 608, and 661;377, 428, 484, 523, 610, and 659;387, 428, 484, 523, 611, and 668;385, 428, 484, 523, 611, and 658;372, 438, 484, 523, 596, and 662; 388, 428, 494, 523, 612, and 667; 369, 439, 484, 523, 613, and 668; 389, 428, 484, 523, 591, and 669; 382, ​​428, 484, 523, 591, and 663; 369, 428, 484, 523, 614, and 659; 381, 428, 484, 523, 591, and 659; 376, 440, 495, 523, 591, and 659; 381, 428, 484, 532, 615, and 662; 381, 42 8, 484, 523, 616, and 670; 390, 441, 484, 523, 591, and 659; 370, 442, 484, 523, 591, and 668; 391, 428, 484, 523, 617, and 671; 377, 428, 484, 533, 595, and 659; 377, 428, 484, 523, 618, and 659; 392, 432, 484, 523, 591, and 659; 377, 443, 484, 534, 619, and 662; 376, 428, 484, 535, 620, and 659; 393, 428, 484, 5 36, 621, and 659; 381, 428, 484, 537, 603, and 671; 383, 444, 489, 523, 622, and 59; 372, 428, 484, 523, 617, and 659; 377, 428, 484, 523, 623, and 659; 376, 428, 484, 538, 607, and 672; 369, 428, 484, 523, 616, and 671; 376, 428, 484, 523, 624, and 659; 394, 428, 484, 523, 591, and 659; 369, 428, 484, 523, 620, and 659; 383, 444, 496, 523, 625, and 660; 385, 428, 484, 523, 602, and 671; 376, 434, 484, 523, 591, and 659; 395, 445, 497, 523, 611, and 663; 396, 446, 487, 523, 626, and 670; 369, 428, 484, 523, 617, and 659; 392, 428, 484, 523, 591, and 662; 394, 428, 484, 523, 606, and 673; 393, 428, 498, 523, 591, and 659;376, 428, 495, 523, 627, and 671; 369, 440, 484, 523, 628, and 671; 397, 428, 484, 539, 591, and 662; 389, 447, 484, 523, 591, and 659; 398, 448, 492, 523, 591, and 670; 373, 449, 490, 523, 611, and 659; 399, 428, 484, 537, 613, and 671; 400, 450, 484, 540, 598, and 659; 401, 428, 484, 541, 591, and 659; 390, 42 8, 484, 542, 629, and 659; 376, 428, 484, 523, 591, and 671; 400, 428, 499, 523, 591, and 659; 381, 428, 484, 523, 630, and 667; 384, 429, 484, 543, 631, and 659; 369, 428, 484, 523, 632, and 664; 402, 436, 484, 523, 597, and 661; 381, 451, 484, 523, 591, and 656; 403, 428, 484, 523, 633, and 659; 369, 429, 484, 5 44, 607, and 659; 404, 429, 484, 523, 630, and 659; 372, 429, 484, 523, 602, and 659; 381, 428, 484, 523, 591, and 665; 388, 428, 484, 523, 610, and 674; 405, 451, 484, 523, 634, and 671; 406, 428, 484, 545, 591, and 661; 400, 428, 484, 523, 635, and 667; 404, 428, 484, 546, 591, and 659; 384, 432, 500, 523, 636, and 664; 376, 452, 501, 547, 591, and 663; 407, 428, 484, 548, 619, and 659; 375, 428, 484, 523, 637, and 659; 372, 428, 484, 523, 607, and 659; 408, 428, 484, 523, 638, and 659; 386, 428, 484, 523, 591, and 659; 369, 428, 497, 523, 591, and 659; 403, 453, 484, 549, 591, and 659; 402, 454, 484, 523, 591, and 659;383, 428, 484, 523, 591, and 659; 383, 434, 500, 523, 591, and 659; 376, 455, 502, 550, 591, and 675; 383, 435, 484, 523, 591, and 664; 369, 428, 484, 523, 602, and 659; 376, 428, 484, 551, 591, and 659; 376, 428, 484, 552, 591, and 659; 383, 428, 484, 553, 591, and 676; 376, 428, 484, 554, 591, and 659; 376, 42 8, 490, 523, 591, and 676; 369, 428, 503, 555, 606, and 659; 409, 428, 484, 523, 591, and 659; 383, 428, 504, 556, 591, and 659; 369, 434, 484, 523, 591, and 659; 390, 428, 484, 523, 637, and 677; 369, 456, 484, 523, 591, and 659; 370, 457, 484, 557, 591, and 659; 383, 428, 484, 558, 591, and 659; 404, 458, 499, 5 23, 591, and 659; 410, 428, 484, 523, 591, and 677; 383, 459, 484, 553, 639, and 659; 407, 428, 489, 549, 591, and 667; 374, 434, 484, 523, 591, and 659; 404, 428, 484, 559, 617, and 666; 411, 460, 505, 523, 591, and 672; 372, 429, 484, 523, 606, and 659; 383, 428, 485, 523, 591, and 659; 386, 460, 490, 523, 601, and 659; 412, 455, 493, 523, 592, and 670; 372, 428, 484, 560, 591, and 667; 372, 428, 484, 523, 606, and 659; 383, 461, 484, 523, 591, and 677; 369, 429, 484, 523, 606, and 659; 369, 434, 484, 530, 591, and 659; 376, 437, 484, 561, 603, and 666; 383, 462, 500, 523, 591, and 659; 383, 428, 484, 562, 591, and 659;400, 463, 484, 523, 616, and 660; 376, 464, 484, 523, 602, and 677; 400, 434, 506, 549, 591, and 659; 369, 436, 507, 523, 591, and 659; 404, 429, 484, 523, 591, and 659; 384, 429, 484, 523, 616, and 659; 369, 429, 484, 523, 591, and 659; 377, 465, 484, 523, 630, and 660; 413, 459, 484, 563, 591, and 659; 369, 45 2, 484, 523, 591, and 663; 414, 466, 484, 523, 640, and 659; 373, 428, 484, 523, 591, and 659; 415, 428, 489, 564, 591, and 659; 369, 428, 484, 523, 591, and 671; 400, 428, 484, 523, 641, and 677; 369, 428, 491, 565, 591, and 662; 403, 458, 508, 523, 642, and 664; 392, 467, 484, 523, 591, and 659; 369, 452, 509, 5 66, 643, and 659; 388, 468, 490, 523, 591, and 659; 369, 452, 484, 523, 644, and 667; 377, 440, 484, 567, 606, and 659; 370, 428, 484, 523, 591, and 656; 384, 467, 487, 523, 591, and 659; 392, 469, 510, 568, 591, and 659; 378, 464, 484, 523, 606, and 659; 377, 428, 484, 569, 645, and 659; 416, 434, 484, 523, 603, and 659; 370, 428, 484, 523, 606, and 659; 370, 428, 511, 570, 628, and 678; 369, 452, 484, 571, 630, and 663; 376, 428, 502, 572, 591, and 664; 417, 428, 484, 523, 591, and 665; 392, 428, 484, 573, 591, and 670; 376, 428, 512, 523, 646, and 659; 377, 428, 484, 574, 591, and 670; 383, 428, 484, 523, 647, and 667;381, 428, 493, 575, 591, and 659; 383, 470, 501, 523, 591, and 659; 404, 471, 484, 537, 591, and 659; 418, 428, 484, 523, 635, and 670; 383, 472, 513, 576, 591, and 659; 41; 9, 428, 484, 577, 606, and 659; 397, 428, 484, 523, 648, and 679; 372, 473, 484, 523, 591, and 664; 383, 474, 484, 523, 592, and 659; 372, 428, 484, 523, 591, and 663; 420, 434, 484, 523, 595, and 659; 370, 428, 514, 523, 591, and 659; 369, 428, 484, 523, 591, and 677; 383, 428, 484, 523, 600, and 659; 369, 475, 484, 523, 591, and 659; 421, 436, 484, 523, 649, and 664; 378, 452, 484, 523, 591, and 659; 376, 476, 515, 578, 641, and 659; 376, 428, 504, 523, 591, and 667; 422, 469, 484, 523, 606, and 660; 383, 461, 484, 523, 616, and 659; 369, 428, 484, 523, 650, and 656; 369, 428, 516, 523, 591, and 659; 405, 460, 484, 52 3, 618, and 659; 413, 431, 484, 523, 591, and 659; 382, ​​428, 484, 579, 621, and 671; 383, 428, 484, 523, 646, and 659; 372, 428, 484, 580, 591, and 660; 406, 429, 484, 527, 591, and 659; 383, 428, 517, 581, 595, and 659; 377, 437, 518, 582, 640, and 664; 369, 477, 487, 583, 651, and 659; 403, 464, 484, 534, 591, and 659; 383, 459, 484, 523, 606, and 659; 369, 454, 484, 584, 591, and 671; 372, 478, 519, 523, 591, and 680; 423, 429, 484, 523, 591, and 681; 383, 428, 484, 585, 591, and 659; 369, 434, 484, 557, 591, and 659; 382, ​​434, 484, 523, 649, and 659; 424, 479, 484, 523, 616, and 660; 376, 429, 484, 586, 591, and 666;425, 428, 484, 523, 592, and 659; 369, 436, 484, 587, 652, and 659; 403, 428, 520, 588, 638, and 659; 417, 460, 484, 523, 591, and 659; 409, 460, 484, 523, 623, and 664; 404, 452, 484, 523, 591, and 659; 369, 428, 484, 523, 591, and 682; 383, 428, 521, 553, 653, and 659; 372, 480, 493, 523, 591, and 659; 369, 428, 484, 523, 654, and 659; 369, 477, 484, 523, 648, and 659; 369, 481, 484, 560, 591, and 659; 369, 452, 484, 523, 591, and 667; 409, 428, 484, 580, 606, and 659; 384, 452, 484, 589, 593, and 661; 377, 482, 484, 523, 591, and 659; 410, 483, 484, 523, 591, and 659; 384, 428, 489, 523, 591, and 659; 426, 429, 484, 523, 622, and 659; 376, 428, 484, 523, 637, and 676; 427, 428, 484, 523, 591, and 671; 373, 452, 506, 523, 591, and 659; 369, 428, 484, 590, 622, and 659; 369, 429, 521, 555, 591, and 661; The following contain the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences as described in 405, 428, 522, 523, 615, and 672; 376, 428, 510, 523, 591, and 659; 384, 428, 484, 523, 626, and 668; 410, 429, 484, 523, 591, and 659; 400, 452, 484, 523, 655, and 659; or 369, 434, 484, 523, 606, and 680.

[0014] In one embodiment, the antibody contains one of the VH amino acid sequences from SEQ ID NOs: 1 to 180.

[0015] In one embodiment, the antibody contains one of the VL amino acid sequences from SEQ ID NOs. 181 to 368.

[0016] In one embodiment, VH and VL are, respectively, sequence numbers 1 and 181;2 and 182;3 and 183;4 and 184;5 and 185;1 and 184;6 and 184;1 and 186;7 and 187;8 and 188;9 and 189;10 and 190;4 and 191;11 and 192;12 and 193;13 and 194;14 and 195;15 and 196;16 and 184;17 and 197;18 and 198;1 and 199;19 and 200;20 and 201;21 and 202;22 and 203;2 3 and 204; 24 and 205; 25 and 184; 4 and 206; 26 and 207; 27 and 208; 28 and 209; 9 and 210; 29 and 211; 30 and 212; 31 and 213; 32 and 214; 33 and 215; 34 and 216; 17 and 217; 1 and 218; 35 and 184; 36 and 184; 35 and 219; 35 and 220; 37 and 184; 38 and 221; 39 and 222; 9 and 223; 9 and 224; 40 and 184; 41 and 225; 8 and 226; 42 and and 227;35 and 228;43 and 229;4 and 230;9 and 231;8 and 232;1 and 233;8 and 234;44 and 184;1 and 235;45 and 236;30 and 237;46 and 184;47 and 238;48 and 239;1 and 230;49 and 190;44 and 240;50 and 184;51 and 241;52 and 242;53 and 243;54 and 184;55 and 244;56 and 245;57 and 246;58 and 247;59 and 248;60 and 249 ;8 and 250;61 and 184;35 and 251;62 and 252;1 and 253;63 and 254;64 and 181;65 and 255;66 and 256;67 and 257;68 and 198;35 and 258;69 and 259;70 and 260;71 and 261;72 and 262;73 and 263;74 and 264;75 and 265;76 and 266;77 and 267;4 and 205;78 and 268;1 and 269;27 and 269;79 and 269;80 and 270;81 and 269;22 and 269;82 and 269;83 and 271;84 and 272;1 and 273;8 and 274;8 and 275;22 and 276;8 and 277;85 and 278;86 and 279;87 and 269;88 and 280;89 and 269;60 and 281;90 and 269;91 and 282;22 and 283;92 and 269;93 and 284;94 and 285;95 and 286;96 and 269;73 and 287;97 and 288;68 and 289;98 and 269;99 and 290;1 00 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304; 72 and 305; 114 and 306; 115 and 307; 116 and 269;117 and 308;118 and 269;110 and 309;119 and 310;120 and 311;121 and 269;122 and 312;123 and 289;9 and 313;124 and 314;120 and 289;125 and 315;110 and 316;126 and 317;127 and 318;49 and 319;128 and 320;9 and 321;22 and 322;129 and 323;130 and 269;131 and 324;132 and 325;133 and 326;134 and 32 7;53 and 328;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340;149 and 341;150 and 342;151 and 343;152 and 344; 94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 356; 166 and 357 ;167 and 358;110 and 334;87 and 359;168 and 360;169 and 269;170 and 269;171 and 269;172 and 361;8 and 362;173 and 304;174 and 269;1 and 363;175 and 364;176 and 365;177 and 269;178 and 366;179 and 269;180 and 367;or containing the amino acid sequences described in 89 and 368.

[0017] In one embodiment, the antibody is a single-stranded variable fragment (scFv).

[0018] In one embodiment, the antibody comprises a heavy chain constant region or its Fc region, which is optionally selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0019] In one embodiment, the heavy chain constant region is human IgG1.

[0020] In one embodiment, the heavy chain constant region is human IgG4.

[0021] In one embodiment, the amino acid sequence of the human IgG 4-chain constant region, numbered according to the EU numbering system, includes P at position 228.

[0022] In one embodiment, the heavy chain constant region is a variant of the wild-type heavy chain constant region, and the variant heavy chain constant region binds to the Fc gamma receptor (FcγR) with lower affinity than the wild-type heavy chain constant region.

[0023] In one embodiment, the amino acid sequence of the heavy chain constant region, in each case, is numbered according to the EU numbering system, with A at position 234, A at position 235, A, Q, or G at position 297, or A or G at position 329.

[0024] In one embodiment, the amino acid sequence of the heavy chain constant region, in each case, is numbered according to the EU numbering system, and includes A at positions 234 and 235, A at positions 234, 235, and 329, or A at positions 234, 235, and G at position 329.

[0025] In one embodiment, the heavy chain constant region is a variant of the wild-type heavy chain constant region, and the variant heavy chain constant region has increased affinity for human neonatal Fc receptor (FcRn) at pH 6 compared to the affinity of the wild-type heavy chain constant region for human FcRn at pH 6.

[0026] In one embodiment, the amino acid sequence of the heavy chain constant region, in each case, is numbered according to the EU numbering system, with L and S at positions 428 and 434, respectively, and K, F, and Y at positions 433, 434, and 436, respectively, or Y, T, and E at positions 252, 254, and 256, respectively.

[0027] In one embodiment, the antibody includes a light chain constant region and, optionally, a human kappa or lambda constant region.

[0028] In one embodiment, the present disclosure provides a polypeptide comprising a VH having one of the CDRH1, CDRH2, and CDRH3 amino acid sequences described in SEQ ID NOs: 1 to 180.

[0029] In one embodiment, the polypeptides are, respectively, SEQ ID NOs: 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381, 428, and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 484; 385, 428, and 484; 372, 438, and 484; 388, 428, and oy 494;369, 439, and 484;389, 428, and 484;381, 428, and 484;376, 440, and 495;390, 441, and 484;370, 442, and 484;391, 428, and 484;392, 432, and 484;377, 443, and 484;393, 428, and 484;383, 444, and 489;394, 428, and 484;383, 444, and 496;376, 434, and 484;395, 445, and 497;396, 446, and 487;392, 4 28, and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484; 402, 436, and 484; 381, 451, and 484; 403, 428, and 484;369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428, and 497; 403, 453, and 484; 402, 454, and 484; 383, 434 , and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428, and 484; 415, 428, and 489; 369, 428, and 491; 403, 458 , and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512; 381, 428, and 493; 383, 470, and 501; 404, 471, and 484;418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515; 376, 428, and 504; 422, 469, and 4 84;369, 428, and 516;405, 460, and 484;413, 431, and 484;406, 429, and 484;383, 428, and 517;377, 437, and 518;369, 477, and 487;403, 464, and 484;369, 454, and 484;372, 478, and 519;423, 429, and 484;382, 434, and 484;424, 479, and and 484;376, 429, and 484;425, 428, and 484;403, 428, and 520;417, 460, and 484;409, 460, and 484;404, 452, and 484;383, 428, and 521;372, 480, and 493;369, 477, and 484;369, 481, and 484;384, 452, and 484;377, 482, and 484;410, 483, and 484; 384, 428, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or containing the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 400, 452, and 484.

[0030] In one embodiment, VH is respectively SEQ ID NOs: 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381 , 428, and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 484; 385, 428, and 484; 372, 438, and 484; 388, 428, and 4 94;369, 439, and 484;389, 428, and 484;381, 428, and 484;376, 440, and 495;390, 441, and 484;370, 442, and 484;391, 428, and 484;392, 432, and 484;377, 443, and 484;393, 428, and 484;383, 444, and 489;394, 428, and 484;383, 444, and 496;376, 434, and 484;395, 445, and 497;396, 446, and 487;392, 42 8, and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484; 402, 436, and 484; 381, 451, and 484; 403, 428, and 484;369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428, and 497; 403, 453, and 484; 402, 454, and 484; 383, 434 , and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428, and 484; 415, 428, and 489; 369, 428, and 491; 403, 458 , and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512; 381, 428, and 493; 383, 470, and 501; 404, 471, and 484;418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515; 376, 428, and 504; 422, 469, and 4 84;369, 428, and 516;405, 460, and 484;413, 431, and 484;406, 429, and 484;383, 428, and 517;377, 437, and 518;369, 477, and 487;403, 464, and 484;369, 454, and 484;372, 478, and 519;423, 429, and 484;382, 434, and 484;424, 479, and and 484;376, 429, and 484;425, 428, and 484;403, 428, and 520;417, 460, and 484;409, 460, and 484;404, 452, and 484;383, 428, and 521;372, 480, and 493;369, 477, and 484;369, 481, and 484;384, 452, and 484;377, 482, and 484;410, 483, and 484; 384, 428, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or containing the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 400, 452, and 484.

[0031] In one embodiment, VH comprises any one of the amino acid sequences described in SEQ ID NOs: 1 to 180.

[0032] In one embodiment, the present disclosure provides a polypeptide comprising a VL containing any one of the CDRL1, CDRL2, and CDRL3 amino acid sequences described in SEQ ID NOs. 181 to 368.

[0033] In one embodiment, polypeptides are, respectively, sequence numbers 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523, 601, and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 661; 523, 610, and 659; 523, 611, and 668; 523, 611, and 658;523, 596, and 662;523, 612, and 667;523, 613, and 668;523, 591, and 669;523, 591, and 663;523, 614, and 659;532, 615, and 662;523, 616, and 670;523, 591, and 668;523, 617, and 671;533, 595, and 659;523, 618, and 659;534, 619, and 662;535, 620, and 659;536, 621, and 659;537, 603, and 671;523, 6 22, and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671; 539, 591, and 662; 523, 591, and 670; 523, 611, and 659;537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610, and 674; 523, 634, and 671; 545, 591, and 661; 523, 635 , and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616, and 660; 523, 602, and 677; 523, 616, and 659; 523, 630 , and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678; 571, 630, and 663; 572, 591, and 664; 573, 591, and 670;523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 523, 595, and 659; 523, 600, and 659; 523, 6 49, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 591, and 659; 584, 591, and 671; 523, 591, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; Containing the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 560, 591, and 659; 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or 523, 606, and 680.

[0034] In one embodiment, VL is, respectively, sequence numbers 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523 , 601, and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 661; 523, 610, and 659; 523, 611, and 668; 523, 611, and 6 58; 523, 596, and 662; 523, 612, and 667; 523, 613, and 668; 523, 591, and 669; 523, 591, and 663; 523, 614, and 659; 532, 615, and 662; 523, 616, and 670; 523, 591, and 668; 523, 617, and 671; 533, 595, and 659; 523, 618, and 659; 534, 619, and 662; 535, 620, and 659; 536, 621, and 659; 537, 603, and 671; 523, 62 2, and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671; 539, 591, and 662; 523, 591, and 670; 523, 611, and 659;537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610, and 674; 523, 634, and 671; 545, 591, and 661; 523, 635 , and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616, and 660; 523, 602, and 677; 523, 616, and 659; 523, 630 , and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678; 571, 630, and 663; 572, 591, and 664; 573, 591, and 670;523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 523, 595, and 659; 523, 600, and 659; 523, 6 49, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 591, and 659; 584, 591, and 671; 523, 591, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; Containing the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 560, 591, and 659; 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or 523, 606, and 680.

[0035] In one embodiment, VL includes any one of the amino acid sequences described in SEQ ID NOs: 181 to 368.

[0036] In one embodiment, the antibodies or polypeptides disclosed herein are conjugated with cytotoxic agents, cell growth inhibitors, toxins, radionuclides, or detectable labels.

[0037] In one embodiment, the present disclosure provides polynucleotides or polypeptides that encode the VH and / or VL of antibodies disclosed herein.

[0038] In one embodiment, the present disclosure provides a vector comprising a polynucleotide disclosed herein.

[0039] In one embodiment, the vector is an adeno-associated virus (AAV) vector.

[0040] In one aspect, this disclosure is, (a) Polynucleotides disclosed herein, (b) vectors disclosed herein, (c) A first polynucleotide encoding the VH or heavy chain of the antibody disclosed herein, and a second polynucleotide encoding the VL or light chain of the antibody disclosed herein, or (d) A recombinant host cell comprising a first vector comprising a first polynucleotide encoding the VH or heavy chain of the antibody disclosed herein, and a second vector comprising a second polynucleotide encoding the VL or light chain of the antibody disclosed herein.

[0041] In one embodiment, the present disclosure provides a pharmaceutical composition comprising an antibody or polypeptide disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, or a host cell disclosed herein, and a pharmaceutically acceptable carrier or excipient.

[0042] In one embodiment, the present disclosure provides a method for producing an antibody or polypeptide, the method comprising culturing a host cell disclosed herein under preferred conditions such that a polynucleotide is expressed and an antibody or polypeptide is produced.

[0043] In one embodiment, the Disclosure provides a method for inhibiting the activity of PC in a subject, the method comprising administering an effective amount of an antibody or polypeptide disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein to the subject.

[0044] In one embodiment, the Disclosure provides a method for treating an inflammatory disorder or degenerative disease in a subject, the method comprising administering an effective amount of an antibody or polypeptide disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein to the subject.

[0045] In one embodiment, the Disclosure provides the use of antibodies or polypeptides, polynucleotides, vectors, host cells, or pharmaceutical compositions disclosed herein for the manufacture of pharmaceuticals for the treatment of inflammatory disorders or degenerative diseases in subjects requiring treatment of such disorders or degenerative diseases.

[0046] In one embodiment, the Disclosure provides antibodies or polypeptides, polynucleotides, vectors, host cells, or pharmaceutical compositions disclosed herein for use in medicine.

[0047] In one embodiment, the Disclosure provides antibodies or polypeptides, polynucleotides, vectors, host cells, or pharmaceutical compositions disclosed herein for use in the treatment of inflammatory disorders or degenerative diseases in subjects requiring treatment of such disorders or degenerative diseases.

[0048] In one embodiment, the inflammatory disorder or degenerative disease is selected from the group consisting of organ reperfusion injury such as myocardial infarction-induced reperfusion injury, Kawasaki disease, non-alcoholic steatohepatitis (NASH), organ transplant rejection, atherosclerosis, type 1 or type 2 diabetes mellitus, rheumatoid arthritis, osteoporosis, acute lung injury, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pain, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), age-related macular degeneration (AMD), stroke, Huntington's disease, frontotemporal dementia (FTD), multiple sclerosis (MS), isolated demyelinating diseases of the central nervous system, osteoarthritis, Crohn's disease, and ulcerative colitis. [Modes for carrying out the invention]

[0049] This disclosure provides polypeptides (e.g., antibodies) that specifically bind to polypeptides (PCs). Also provided are pharmaceutical compositions comprising these polypeptides, nucleic acids encoding these polypeptides, expression vectors and host cells for producing these polypeptides, and methods for treating a subject using these antibodies. The polypeptides provided herein are particularly advantageous because they bind to PCs with high affinity, exhibit good stability, stress tolerance, and immunogenicity profiles, and can be produced in high yield. The polypeptides provided herein are particularly useful for treating inflammatory disorders or degenerative diseases in a subject.

[0050] definition As used herein, the terms “antibody” and “antibodies” include full-length antibodies, antigen-binding fragments of full-length antibodies, and molecules containing antibody CDRs, VH regions, and / or VL regions. Examples of antibodies include, but are not limited to, monoclonal antibodies, recombinant antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies containing two heavy chains and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain-antibody heavy chain pairs, intrabodies, heteroconjugate antibodies, antibody-drug conjugates, single-domain antibodies, monovalent antibodies, single-chain antibodies or single-chain Fvs(scFv), camelized antibodies, aphibodies, Fab fragments, F(ab')2 fragments, disulfide-linked Fvs(sdFv), anti-idiotype (anti-Id) antibodies (e.g., anti-anti-Id antibodies), and any of the antigen-binding fragments described above. In certain embodiments, the antibodies described herein refer to a population of polyclonal antibodies. The antibody may be any type of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a or IgG2b). In certain embodiments, the antibody described herein is an IgG antibody, or a class thereof (e.g., human IgG1 or IgG4) or a subclass thereof. In specific embodiments, the antibody is a humanized monoclonal antibody. In another specific embodiment, the antibody is a human monoclonal antibody.

[0051] A "multispecific antibody" is an antibody that specifically binds to two or more different antigens, or to two or more different regions of the same antigen (e.g., a bispecific antibody). Multispecific antibodies include bispecific antibodies that contain two different antigen-binding sites (excluding the Fc region). Examples of multispecific antibodies include recombinant antibodies, human antibodies, humanized antibodies, resurfaced antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies containing two heavy chains and two light chain molecules, antibody light chain monomers, heteroconjugate antibodies, linked single-chain antibodies or linked single-chain Fvs (scFv), camelized antibodies, aphibodies, linked Fab fragments, F(ab')2 fragments, chemically linked Fv, and disulfide-linked Fvs (sdFv). A multispecific antibody can be any type of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a or IgG2b). In certain embodiments, the multispecific antibody described herein is an IgG antibody, or a class thereof (e.g., human IgG1, IgG2, or IgG4) or a subclass thereof.

[0052] As used herein, the terms “CDR” or “complementarity-determining region” refer to non-adjacent antigen-binding sites found within the variable regions of heavy and light chain polypeptides. These specific regions are described, for example, in Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977), and Kabat et al., Sequences of protein of immunological interest. (1991), Chothia et al., J. Mol. Biol. 196:901-917 (1987), and MacCallum et al., J. Mol. Biol. 262:732-745 (1996), all of which are incorporated herein by reference, and the definition herein includes overlaps or subsets of amino acid residues when compared to one another. In certain embodiments, the term "CDR" refers to the CDR as defined by MacCallum et al., J.Mol.Biol.262:732-745 (1996), and Martin A. "Protein Sequence and Structure Analysis of Antibody Variable Domains," Antibody Engineering, Kontermann and Duebel, eds., Chapter 31, pp.422-439, Springer-Verlag, Berlin (2001). In certain embodiments, the term "CDR" refers to the CDR as defined by Kabat et al., J.Biol.Chem.252, 6609-6616 (1977), and Kabat et al., Sequences of protein of immunological interest. (1991). In certain embodiments, the heavy chain CDR and light chain CDR of an antibody are defined using different conventions. In certain embodiments, heavy chain CDRs and / or light chain CDRs are defined by performing a structural analysis of the antibody and identifying residues in the variable region(s) predicted to contact the epitope region of the target molecule (e.g., PC). CDRH1, CDRH2, and CDRH3 represent heavy chain CDRs, and CDRL1, CDRL2, and CDRL3 represent light chain CDRs.

[0053] As used herein, the terms “variable region” and “variable domain” are used interchangeably and are common in the art. A variable region refers to a portion of an antibody, generally a portion of the light chain or heavy chain, typically the nearly amino-terminus 110–120 or 110–125 amino acids in the mature heavy chain, and about 90–115 amino acids in the mature light chain, where the sequence differs significantly between antibodies and is used in the binding and specificity of a particular antibody to a particular antigen. Sequence variability is concentrated in those regions called complementarity-determining regions (CDRs), while more highly conserved regions within the variable region are called framework regions (FRs). While we do not wish to be constrained by any particular mechanism or theory, the CDRs of the light and heavy chains are thought to be primarily involved in antibody-antigen interaction and specificity. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region includes rodent or mouse CDRs and human framework regions (FRs). In certain embodiments, the variable region includes a rodent or mouse CDR and a primate (e.g., non-human primate) framework region (FR).

[0054] As used herein, the terms "VH" and "VL" refer to the heavy chain and light chain variable regions of an antibody, respectively, as described in Kabat et al., (1991) Sequences of Proteins of Immunological Interest (NIH Publication No. 91-3242, Bethesda), and are incorporated herein by reference in their entirety.

[0055] As used herein, the term “constant region” is common in the art. The constant region is the carboxyl-terminal portion of the light and / or heavy chain that does not directly participate in the binding of the antibody to an antigen, for example, but can exhibit various effector functions, such as interaction with Fc receptors (e.g., Fc gamma receptors).

[0056] As used herein, the term “heavy chain,” when used in reference to an antibody, can refer to any different type based on the amino acid sequence of the constant region, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), which respectively give rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes, including subclasses of IgG, e.g., IgG1, IgG2, IgG3, and IgG4.

[0057] As used herein, the term “light chain,” when used in reference to an antibody, may refer to any different type, for example, kappa (κ) or lambda (λ), based on the amino acid sequence of the constant region. Light chain amino acid sequences are well known in the art. In specific embodiments, the light chain is a human light chain.

[0058] As used herein, the terms “specifically bind,” “specifically recognize,” “immunospecifically bind,” and “immunospecifically recognize” are analogous terms in the context of antibodies and refer to molecules that bind to an antigen (e.g., an epitope or immune complex) in such a way that such binding is understood by those skilled in the art. For example, a molecule that specifically binds to an antigen may bind to other peptides or polypeptides with generally lower affinity, as measured by immunoassays, BIAcore®, KinExA 3000 instrument (Sapidyne Instruments, Boise, ID), or other assays known in the art. In specific embodiments, a molecule that specifically binds to an antigen may be referred to as K if the molecule binds nonspecifically to another antigen. A K is at least 2 log (e.g., 10 times), 2.5 log, 3 log, or 4 log greater. A It then binds to the antigen.

[0059] As used herein, the term “affinity” refers to the sum of the non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, “binding affinity” refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y can generally be expressed by a dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein.

[0060] As used herein, the term “EU numbering system” refers to the EU numbering convention for constant regions of antibodies, as described in Edelman, GM et al., Proc. Natl. Acad. USA, 63, 78-85 (1969), and Kabat et al., Sequences of Proteins of Immunological Interest, USDept. Health and Human Services, 5th edition, 1991, each of which is incorporated herein in whole by reference.

[0061] As used herein, the terms “to treat,” “to treat,” and “treatment” refer to the therapeutic or preventive measures described herein. A “treatment” method involves administering antibodies to a subject who has or is prone to having a disease or disorder in order to prevent, cure, delay, reduce the severity of, or improve one or more symptoms of a disease or disorder, or a recurrent disease or disorder, or to extend the survival of the subject beyond the survival expected in the absence of such treatment.

[0062] As used herein, in the context of therapy, the term “effective dose” refers to the amount of therapy that achieves the desired preventive or therapeutic effect.

[0063] As used herein, the term “Subject” includes any human or non-human animal. In certain embodiments, the subject is a human or a non-human mammal. In certain embodiments, the subject is a human.

[0064] Where used herein with respect to antibodies or polynucleotides, the term “isolated” means an antibody or polynucleotide isolated from one or more contaminants present in the natural source of the antibody or polynucleotide (e.g., polypeptides, polynucleotides, lipids, or carbohydrates). All examples of “isolated antibodies” described herein are intended to be antibodies that may be further isolated but do not require isolation. All examples of “isolated polynucleotides” described herein are intended to be polynucleotides that may be further isolated but do not require isolation. All examples of “antibodies” described herein are intended to be antibodies that may be further isolated but do not require isolation. All examples of “polynucleotides” described herein are intended to be polynucleotides that may be further isolated but do not require isolation.

[0065] The determination of the "percentage of identity" between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be achieved using mathematical algorithms. Specific non-restrictive examples of mathematical algorithms used for comparing two sequences are those of Karlin S & Altschul SF (1990) PNAS 87:2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90:5873-5877, each of which is incorporated herein by reference in its entirety. Such algorithms are incorporated into the NBLAST and XBLAST programs of Altschul SF et al., (1990) J Mol Biol 215:403, which is also incorporated herein by reference in its entirety. To obtain nucleotide sequences homologous to the nucleic acid molecules described herein, a BLAST nucleotide search can be performed using, for example, an NBLAST nucleotide program parameter set such as score = 100, word length = 12. To obtain amino acid sequences homologous to the protein molecules described herein, a BLAST protein search can be performed using, for example, an XBLAST program parameter set such as score 50 and word length = 3. To obtain gap alignment for comparison, Gapped BLAST can be used, as described by Altschul SF et al., (1997) Nuc Acids Res 25:3389-3402. Alternatively, PSI BLAST can be used to perform iterative searches to detect intermolecular distance relationships (ibid.). When using the BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used (see, for example, the National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov).Another specific, non-restrictive example of a mathematical algorithm used for sequence comparison is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is incorporated herein by reference in its entirety. Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When using the ALIGN program to compare amino acid sequences, the PAM120 weight residue table, gap length penalty 12, and gap penalty 4 can be used.

[0066] The percentage of identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. When calculating the percentage of identity, typically only exact matches are counted.

[0067] anti-PC antibody In one embodiment, the present disclosure provides a polypeptide (e.g., an antibody) that specifically binds to PC. Table 1 shows the amino acid sequence of an exemplary antibody. [Table 1] TIFF2026509512000002.tif253170TIFF2026509512000003.tif252170TIFF20265095120 00004.tif252170TIFF2026509512000005.tif253170TIFF2026509512000006.tif173170

[0068] In certain embodiments, the Disclosure provides an antibody that specifically binds to PC, the antibody comprising a VH domain containing one, two, or all three of the VH domain CDRs listed in Table 1. In certain embodiments, the antibody comprises CDRH1 of the VH domain listed in Table 1. In certain embodiments, the antibody comprises CDRH2 of the VH domain listed in Table 1. In certain embodiments, the antibody comprises CDRH3 of the VH domain listed in Table 1.

[0069] In certain embodiments, the Disclosure provides an antibody that specifically binds to PC, the antibody comprising a VL domain containing one, two, or all three of the VL domain CDRs disclosed in Table 1. In certain embodiments, the antibody comprises CDRL1 of the VL domains listed in Table 1. In certain embodiments, the antibody comprises CDRL2 of the VL domains listed in Table 1. In certain embodiments, the antibody comprises CDRL3 of the VL domains listed in Table 1.

[0070] The individual CDRs of the antibodies disclosed herein can be determined according to any CDR numbering scheme known in the art.

[0071] In certain embodiments, one or more CDRs of the antibodies disclosed herein can be determined according to Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest (1991), each of which is incorporated herein by reference in whole.

[0072] In certain embodiments, the Disclosure provides antibodies that specifically bind to PC, the antibodies comprising CDRs of the antibodies disclosed in Table 1 herein, as determined by the Kabat numbering scheme.

[0073] In certain embodiments, one or more CDRs of the antibodies disclosed herein can be determined according to the Chothia numbering scheme that points to the position of the immunoglobulin structural loop (see, for example, Chothia C & Lesk AM, (1987), J Mol Biol 196:901-917; Al-Lazikani B et al., (1997), J Mol Biol 273:927-948; Chothia C et al., (1992), J Mol Biol 227:799-817; Tramontano A et al., (1990), J Mol Biol 215(1):175-82; and U.S. Patent No. 7,709,226, all of which are incorporated herein by reference in their entirety).

[0074] In certain embodiments, the Disclosure provides antibodies that specifically bind to PC, the antibodies comprising the CDRs of the antibodies disclosed in Table 1 of this Specification, as determined by the Chothia numbering system.

[0075] In certain embodiments, one or more CDRs of the antibodies disclosed herein can be determined according to MacCallum RM et al., (1996) J Mol Biol 262:732-745, the whole thereof, which is incorporated herein by reference. See also, for example, Martin A. “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Duebel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001), the whole thereof, which is incorporated herein by reference.

[0076] In certain embodiments, the Disclosure provides antibodies that specifically bind to PC, the antibodies comprising the CDRs of the antibodies disclosed in Table 1 of this Specification, as determined by the MacCallum numbering system.

[0077] In certain embodiments, the CDR of the antibodies disclosed herein can be determined according to the IMGT numbering system described in Lefranc MP, (1999) The Immunologist 7:132-136, Lefranc MP et al., (1999) Nucleic Acids Res 27:209-212 (each of which is incorporated herein by reference in whole), and Lefranc MP et al., (2009) Nucleic Acids Res 37:D1006-D1012.

[0078] In certain embodiments, the Disclosure provides antibodies that specifically bind to PC, the antibodies comprising the CDRs of the antibodies disclosed in Table 1 herein, as determined by the IMGT numbering system.

[0079] In certain embodiments, the CDR of the antibodies disclosed herein can be determined according to an AbM numbering scheme, which refers to the AbM hypervariable region, represents the compromise between the Kabat CDR and the Chothia structural loop, and is used by Oxford Molecular's AbM antibody modeling software (Oxford Molecular Group, Inc.), the entire scheme of which is incorporated herein by reference.

[0080] In certain embodiments, the Disclosure provides antibodies that specifically bind to PC, the antibodies comprising CDRs of the antibodies disclosed in Table 1 of this Specification, as determined by an AbM numbering scheme.

[0081] In certain embodiments, the CDR of the antibodies disclosed herein can be determined according to the AHo numbering system described in Honegger and Plueckthun, A., J. Mol. Biol. 309:657-670 (2001).

[0082] In certain embodiments, the Disclosure provides antibodies that specifically bind to PC, the antibodies comprising the CDRs of the antibodies disclosed in Table 1 herein, as determined by the AHo numbering system.

[0083] In certain embodiments, the individual CDRs of the antibodies disclosed herein are determined independently, either according to one of the numbering schemes of Kabat, Chothia, MacCallum, IMGT, AHo, or AbM, or by molecular structural analysis, which identifies residues within the variable region(s) predicted to contact the epitope region of the PC.

[0084] In certain embodiments, the Disclosure provides antibodies or polypeptides that specifically bind to PCs comprising a VH comprising one of the CDRH1, CDRH2, and CDRH3 amino acid sequences from the VH amino acid sequences described in SEQ ID NOs. 1 to 180, and a VL comprising one of the CDRL1, CDRL2, and CDRL3 amino acid sequences from the VL amino acid sequences described in SEQ ID NOs. 181 to 368, each CDR independently determined according to one of the Kabat, Chothia, MacCallum, IMGT, AHo, or AbM numbering schemes, or by structural analysis of the molecule, where the structural analysis identifies residues in a variable region(s) predicted to contact the epitope region of the PC.

[0085] In certain embodiments, the Disclosure provides antibodies or polypeptides that specifically bind to PC, the antibodies or polypeptides being, respectively, SEQ ID NOs: 1 and 181; 2 and 182; 3 and 183; 4 and 184; 5 and 185; 1 and 184; 6 and 184; 1 and 186; 7 and 187; 8 and 188; 9 and 189; 10 and 190; 4 and 191; 11 and 192; 12 and 193; 13 and 194; 14 and 195; 15 and 196; 16 and 184; 17 and 197; 18 and 198; 1 and 199; 19 and 200; 20 and 201; 21 and 202; 22 and 203; 23 and 204; 24 and 205; 25 and 184; 4 and 206; 26 and 207; 27 and 208; 28 and 209; 9 and 210; 29 and 211; 30 and 212; 31 and 213; 32 and 214; 33 and 215; 34 and 216; 17 and 217; 1 and 218; 35 and 184; 36 and 184; 35 and 219; 35 and 220; 37 and 184; 38 and 221; 39 and 222; 9 and 223; 9 and 224;40 and 184;41 and 225;8 and 226;42 and 227;35 and 228;43 and 229;4 and 230;9 and 231;8 and 232;1 and 233;8 and 234;44 and 184;1 and 235;45 and 236;30 and 237;46 and 184;47 and 238;48 and 239;1 and 230;49 and 190;44 and 240;50 and 184;51 and 241;52 and 242;53 and 243;54 and 184;55 and 244;56 and 24 5;57 and 246;58 and 247;59 and 248;60 and 249;8 and 250;61 and 184;35 and 251;62 and 252;1 and 253;63 and 254;64 and 181;65 and 255;66 and 256;67 and 257;68 and 198;35 and 258;69 and 259;70 and 260;71 and 261;72 and 262;73 and 263;74 and 264;75 and 265;76 and 266;77 and 267;4 and 205;78 and 268;1 and 269;27 and 269; 79 and 269; 80 and 270; 81 and 269; 22 and 269; 82 and 269; 83 and 271; 84 and 272; 1 and 273; 8 and 274; 8 and 275; 22 and 276; 8 and 277; 85 and 278; 86 and 279; 87 and 269; 88 and 280; 89 and 269; 60 and 281; 90 and 269; 91 and 282; 22 and 283; 92 and 269; 93 and 284; 94 and 285; 95 and 286; 96 and 269; 73 and 287; 97 and 288; 68 and 289; 98 and 269; 99 and 290; 100 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304;72 and 305;114 and 306;115 and 307;116 and 269;117 and 308;118 and 269;110 and 309;119 and 310;120 and 311;121 and 269;122 and 312;123 and 289;9 and 313;124 and 314;120 and 289;125 and 315;110 and 316;126 and 317;127 and 318;49 and 319;128 and 320;9 and 321;22 and 322;129 and 323;130 and 269 ;131 and 324;132 and 325;133 and 326;134 and 327;53 and 328;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340; 149 and 341; 150 and 342; 151 and 343; 152 and 344; 94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 356; 166 and 357; 167 and The VH and VL amino acid sequences described in 358;110 and 334;87 and 359;168 and 360;169 and 269;170 and 269;171 and 269;172 and 361;8 and 362;173 and 304;174 and 269;1 and 363;175 and 364;176 and 365;177 and 269;178 and 366;179 and 269;180 and 367; or 89 and 368 include the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences.

[0086] In certain embodiments, the Disclosure provides antibodies or polypeptides that specifically bind to PC, the antibodies or polypeptides being, respectively, SEQ ID NOs: 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381, 428, and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 48 4;385, 428, and 484;372, 438, and 484;388, 428, and 494;369, 439, and 484;389, 428, and 484;381, 428, and 484;376, 440, and 495;390, 441, and 484;370, 442, and 484;391, 428, and 484;392, 432, and 484;377, 443, and 484;393, 428, and 484;383, 444, and 489;394, 428, and 484;383, 444, and 496;376, 43 4, and 484; 395, 445, and 497; 396, 446, and 487; 392, 428, and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484;402, 436, and 484; 381, 451, and 484; 403, 428, and 484; 369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428 , and 497; 403, 453, and 484; 402, 454, and 484; 383, 434, and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428 , and 484; 415, 428, and 489; 369, 428, and 491; 403, 458, and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512;381, 428, and 493; 383, 470, and 501; 404, 471, and 484; 418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515;376, 428, and 504;422, 469, and 484;369, 428, and 516;405, 460, and 484;413, 431, and 484;406, 429, and 484;383, 428, and 517;377, 437, and 518;369, 477, and 487;403, 464, and 484;369, 454, and 484;372, 478, and 519;423, 429, and 484;382, 4 34, and 484; 424, 479, and 484; 376, 429, and 484; 425, 428, and 484; 403, 428, and 520; 417, 460, and 484; 409, 460, and 484; 404, 452, and 484; 383, 428, and 521; 372, 480, and 493; 369, 477, and 484; 369, 481, and 484; 384, 452, and 484; 377, 482, and 484; VH containing the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 410, 483, and 484; 384, 428, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or 400, 452, and 484.

[0087] In certain embodiments, the Disclosure provides antibodies or polypeptides that specifically bind to PC, the antibodies or polypeptides being, respectively, SEQ ID NOs: 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523, 601, and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 66 1; 523, 610, and 659; 523, 611, and 668; 523, 611, and 658; 523, 596, and 662; 523, 612, and 667; 523, 613, and 668; 523, 591, and 669; 523, 591, and 663; 523, 614, and 659; 532, 615, and 662; 523, 616, and 670; 523, 591, and 668; 523, 617, and 671; 533, 595, and 659; 523, 618, and 659; 534, 619, and 662; 535, 62 0, and 659; 536, 621, and 659; 537, 603, and 671; 523, 622, and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671;539, 591, and 662; 523, 591, and 670; 523, 611, and 659; 537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610 , and 674; 523, 634, and 671; 545, 591, and 661; 523, 635, and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616 , and 660; 523, 602, and 677; 523, 616, and 659; 523, 630, and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678;571, 630, and 663; 572, 591, and 664; 573, 591, and 670; 523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 52 3, 595, and 659; 523, 600, and 659; 523, 649, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 5 91, and 659; 584, 591, and 671; 523, 591, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; 560, 591, and 659; 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or VL containing the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 523, 606, and 680.

[0088] In certain embodiments, the Disclosure provides an antibody or polypeptide that specifically binds to PC, the antibody or polypeptide comprising a VH including CDRH1, CDRH2, and CDRH3 regions and a VL including CDRL1, CDRL2, and CDRL3 regions, where the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions are respectively SEQ ID NOs: 369, 428, 484, 523, 591, and 656; 370, 429, 485, 523, 592, and 657; 371, 428, 486, 523, 593, and 658;372, 428, 484, 523, 591, and 659;373, 429, 484, 523, 594, and 659;369, 428, 484, 523, 591, and 659;374, 430, 484, 523, 591, and 659;369, 428, 484, 523, 595, and 660;375, 428, 487, 523, 596, and 659;376, 428, 484, 524, 591, and 659;377, 428, 484, 523, 591, and 661;378, 428, 484, 523, 591, and 6 62;372, 428, 484, 523, 597, and 658;379, 428, 484, 523, 598, and 662;372, 428, 488, 525, 599, and 659;376, 431, 484, 523, 600, and 663;380, 432, 484, 523, 591, and 664;372, 428, 489, 523, 601, and 659;381, 428, 490, 523, 591, and 659;382, 428, 484, 526, 597, and 665;381, 428, 491, 523, 602, and 65 9;369, 428, 484, 527, 591, and 661;377, 428, 492, 528, 591, and 662;376, 433, 484, 529, 603, and 662;383, 434, 484, 523, 604, and 666;383, 428, 484, 523, 605, and 667;384, 435, 484, 530, 606, and 659;369, 436, 484, 523, 607, and 659;372, 437, 487, 523, 591, and 659;372, 428, 484, 523, 608, and 659;385, 428, 493, 523, 609, and 662; 386, 428, 484, 531, 591, and 663; 372, 437, 484, 523, 608, and 661; 377, 428, 484, 523, 610, and 659; 387, 428, 484, 523, 611, and 668; 385, 428, 484, 523, 611, and 658; 372, 438, 484, 523, 596, and 662; 388, 428, 494, 523, 612, and 667; 369, 439, 484, 523, 613, and 668; 389, 42 8, 484, 523, 591, and 669; 382, ​​428, 484, 523, 591, and 663; 369, 428, 484, 523, 614, and 659; 381, 428, 484, 523, 591, and 659; 376, 440, 495, 523, 591, and 659; 381, 428, 484, 532, 615, and 662; 381, 428, 484, 523, 616, and 670; 390, 441, 484, 523, 591, and 659; 370, 442, 484, 523, 591, and 668; 391, 428, 484, 5 23, 617, and 671; 377, 428, 484, 533, 595, and 659; 377, 428, 484, 523, 618, and 659; 392, 432, 484, 523, 591, and 659; 377, 443, 484, 534, 619, and 662; 376, 428, 484, 535, 620, and 659; 393, 428, 484, 536, 621, and 659; 381, 428, 484, 537, 603, and 671; 383, 444, 489, 523, 622, and 59; 372, 428, 484, 523, 617, and 659; 377, 428, 484, 523, 623, and 659; 376, 428, 484, 538, 607, and 672; 369, 428, 484, 523, 616, and 671; 376, 428, 484, 523, 624, and 659; 394, 428, 484, 523, 591, and 659; 369, 428, 484, 523, 620, and 659; 383, 444, 496, 523, 625, and 660; 385, 428, 484, 523, 602, and 671; 376, 434, 484, 523, 591, and 659;395, 445, 497, 523, 611, and 663; 396, 446, 487, 523, 626, and 670; 369, 428, 484, 523, 617, and 659; 392, 428, 484, 523, 591, and 662; 394, 428, 484, 523, 606, and 673; 393, 428, 498, 523, 591, and 659; 376, 428, 495, 523, 627, and 671; 369, 440, 484, 523, 628, and 671; 397, 428, 484, 539, 591, and 662; 389, 44 7, 484, 523, 591, and 659; 398, 448, 492, 523, 591, and 670; 373, 449, 490, 523, 611, and 659; 399, 428, 484, 537, 613, and 671; 400, 450, 484, 540, 598, and 659; 401, 428, 484, 541, 591, and 659; 390, 428, 484, 542, 629, and 659; 376, 428, 484, 523, 591, and 671; 400, 428, 499, 523, 591, and 659; 381, 428, 484, 5 23, 630, and 667; 384, 429, 484, 543, 631, and 659; 369, 428, 484, 523, 632, and 664; 402, 436, 484, 523, 597, and 661; 381, 451, 484, 523, 591, and 656; 403, 428, 484, 523, 633, and 659; 369, 429, 484, 544, 607, and 659; 404, 429, 484, 523, 630, and 659; 372, 429, 484, 523, 602, and 659; 381, 428, 484, 523, 591, and 665; 388, 428, 484, 523, 610, and 674; 405, 451, 484, 523, 634, and 671; 406, 428, 484, 545, 591, and 661; 400, 428, 484, 523, 635, and 667; 404, 428, 484, 546, 591, and 659; 384, 432, 500, 523, 636, and 664; 376, 452, 501, 547, 591, and 663; 407, 428, 484, 548, 619, and 659; 375, 428, 484, 523, 637, and 659;372, 428, 484, 523, 607, and 659; 408, 428, 484, 523, 638, and 659; 386, 428, 484, 523, 591, and 659; 369, 428, 497, 523, 591, and 659; 403, 453, 484, 549, 591, and 659; 402, 454, 484, 523, 591, and 659; 383, 428, 484, 523, 591, and 659; 383, 434, 500, 523, 591, and 659; 376, 455, 502, 550, 591, and 675; 383, 43 5, 484, 523, 591, and 664; 369, 428, 484, 523, 602, and 659; 376, 428, 484, 551, 591, and 659; 376, 428, 484, 552, 591, and 659; 383, 428, 484, 553, 591, and 676; 376, 428, 484, 554, 591, and 659; 376, 428, 490, 523, 591, and 676; 369, 428, 503, 555, 606, and 659; 409, 428, 484, 523, 591, and 659; 383, 428, 504, 5 56, 591, and 659; 369, 434, 484, 523, 591, and 659; 390, 428, 484, 523, 637, and 677; 369, 456, 484, 523, 591, and 659; 370, 457, 484, 557, 591, and 659; 383, 428, 484, 558, 591, and 659; 404, 458, 499, 523, 591, and 659; 410, 428, 484, 523, 591, and 677; 383, 459, 484, 553, 639, and 659; 407, 428, 489, 549, 591, and 667; 374, 434, 484, 523, 591, and 659; 404, 428, 484, 559, 617, and 666; 411, 460, 505, 523, 591, and 672; 372, 429, 484, 523, 606, and 659; 383, 428, 485, 523, 591, and 659; 386, 460, 490, 523, 601, and 659; 412, 455, 493, 523, 592, and 670; 372, 428, 484, 560, 591, and 667; 372, 428, 484, 523, 606, and 659;383, 461, 484, 523, 591, and 677; 369, 429, 484, 523, 606, and 659; 369, 434, 484, 530, 591, and 659; 376, 437, 484, 561, 603, and 666; 383, 462, 500, 523, 591, and 659; 383, 428, 484, 562, 591, and 659; 400, 463, 484, 523, 616, and 660; 376, 464, 484, 523, 602, and 677; 400, 434, 506, 549, 591, and 659; 369, 43 6, 507, 523, 591, and 659; 404, 429, 484, 523, 591, and 659; 384, 429, 484, 523, 616, and 659; 369, 429, 484, 523, 591, and 659; 377, 465, 484, 523, 630, and 660; 413, 459, 484, 563, 591, and 659; 369, 452, 484, 523, 591, and 663; 414, 466, 484, 523, 640, and 659; 373, 428, 484, 523, 591, and 659; 415, 428, 489, 5 64, 591, and 659; 369, 428, 484, 523, 591, and 671; 400, 428, 484, 523, 641, and 677; 369, 428, 491, 565, 591, and 662; 403, 458, 508, 523, 642, and 664; 392, 467, 484, 523, 591, and 659; 369, 452, 509, 566, 643, and 659; 388, 468, 490, 523, 591, and 659; 369, 452, 484, 523, 644, and 667; 377, 440, 484, 567, 606, and 659; 370, 428, 484, 523, 591, and 656; 384, 467, 487, 523, 591, and 659; 392, 469, 510, 568, 591, and 659; 378, 464, 484, 523, 606, and 659; 377, 428, 484, 569, 645, and 659; 416, 434, 484, 523, 603, and 659; 370, 428, 484, 523, 606, and 659; 370, 428, 511, 570, 628, and 678; 369, 452, 484, 571, 630, and 663;376, 428, 502, 572, 591, and 664; 417, 428, 484, 523, 591, and 665; 392, 428, 484, 573, 591, and 670; 376, 428, 512, 523, 646, and 659; 377, 428, 484, 574, 591, and 670; 383, 428, 484, 5; 23, 647, and 667; 381, 428, 493, 575, 591, and 659; 383, 470, 501, 523, 591, and 659; 404, 471, 484, 537, 591, and 659; 418, 428, 484, 523, 635, and 670; 383, 472, 513, 576, 591, and 659; 419, 428, 484, 577, 606, and 659; 397, 428, 484, 523, 648, and 679; 372, 473, 484, 523, 591, and 664; 383, 474, 484, 523, 592, and 659; 372, 428, 484, 523, 591, and 663; 420, 434, 484, 523, 595, and 659; 370, 428, 514, 523, 591, and 659; 369, 428, 484, 523, 591, and 677; 383, 428, 484, 523, 600, and 659; 369, 475, 484, 523, 591, and 659; 421, 436, 484, 523, 649, and 664; 378, 452, 484, 523, 591, and 659; 376, 476, 515, 578, 641, and 659; 376, 428, 504, 523, 591, and 667; 422, 469, 484, 523, 606, and 660; 383, 461, 484, 523, 616, and 659; 369, 428, 484, 523, 650, and 656; 369, 428, 516, 523, 591, and 659; 405, 460, 484, 523, 618, and 659; 413, 431, 484, 523, 591, and 659; 382, ​​428, 484, 579, 621, and 671; 383, 428, 484, 523, 646, and 659; 372, 428, 484, 580, 591, and 660; 406, 429, 484, 527, 591, and 659; 383, 428, 517, 581, 595, and 659; 377, 437, 518, 582, 640, and 664; 369, 477, 487, 583, 651, and 659; 403, 464, 484, 534, 591, and 659; 383, 459, 484, 523, 606, and 659; 369, 454, 484, 584, 591, and 671; 372, 478, 519, 523, 591, and 680;423, 429, 484, 523, 591, and 681; 383, 428, 484, 585, 591, and 659; 369, 434, 484, 557, 591, and 659; 382, ​​434, 484, 523, 649, and 659; 424, 479, 484, 523, 616, and 660; 376, 429, 484, 586, 591, and 666; 425, 428, 484, 523, 592, and 659; 369, 436, 484, 587, 652, and 659; 403, 428, 520, 588, 638, and 659; 417, 460, 484, 523, 591, and 659; 409, 460, 484, 523, 623, and 664; 404, 452, 484, 523, 591, and 659; 369, 428, 484, 523, 591, and 682; 383, 428, 521, 553, 653, and 659; 372, 480, 493, 523, 591, and 659; 369, 428, 484, 523, 654, and 659; 369, 477, 484, 523, 648, and 659; 369, 481, 484, 560, 591, and 659; 369, 452, 484, 523, 591, and 667; 409, 428, 484, 580, 606, and 659; 384, 452, 484, 589, 593, and 661; 377, 482, 484, 523, 591, and 659; 410, 483, 484, 523, 591, and 659; 384, 428, 489, 523, 591, and 659; 426, 429, 484, 523, 622, and 659; 376, 428, 484, 523, 637, and 676; 427, 428, 484, 523, 591, and 671; 373, 452, 506, Includes the amino acid sequences described in 523, 591, and 659; 369, 428, 484, 590, 622, and 659; 369, 429, 521, 555, 591, and 661; 405, 428, 522, 523, 615, and 672; 376, 428, 510, 523, 591, and 659; 384, 428, 484, 523, 626, and 668; 410, 429, 484, 523, 591, and 659; 400, 452, 484, 523, 655, and 659; or 369, 434, 484, 523, 606, and 680.

[0089] In certain embodiments, the disclosure provides an antibody or polypeptide that specifically binds to PC, comprising VH having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to any one of the amino acid sequences described in SEQ ID NOs: 1 to 180. amino acid sequence of VH consists of any one of the amino acid sequences described in SEQ ID NOs: 1 to 180.

[0090] In certain embodiments, the Disclosure provides an antibody or polypeptide that specifically binds to PC, comprising a VL containing an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to any one of the amino acid sequences described in SEQ ID NOs. 181-368. 201 embodiments. The Disclosure provides an antibody or polypeptide that specifically binds to PC, comprising a VL containing any one of the amino acid sequences described in SEQ ID NOs. 181-368. 301 embodiments. The amino acid sequence of the VL consists of any one of the amino acid sequences described in SEQ ID NOs. 181-368.

[0091] In certain embodiments, the Disclosure provides an antibody or polypeptide that specifically binds to PC, the antibody comprising VH having an amino acid sequence identical by at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) to any one of the amino acid sequences described in SEQ ID NOs. 181-368, the VL having an amino acid sequence identical by at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) to any one of the amino acid sequences described in SEQ ID NOs. 181-368. In certain embodiments, the disclosure provides an antibody or polypeptide that specifically binds to PC, comprising VH containing any one of the amino acid sequences described in SEQ ID NOs: 1 to 180, and VL containing any one of the amino acid sequences described in SEQ ID NOs: 181 to 368. In certain embodiments, the amino acid sequence of VH consists of any one of the amino acid sequences described in SEQ ID NOs: 1 to 180, and the amino acid sequence of VL consists of any one of the amino acid sequences described in SEQ ID NOs: 181 to 368.

[0092] In certain embodiments, the Disclosure provides antibodies or polypeptides that specifically bind to PC, the antibodies being, respectively, SEQ ID NOs: 1 and 181;2 and 182;3 and 183;4 and 184;5 and 185;1 and 184;6 and 184;1 and 186;7 and 187;8 and 188;9 and 189;10 and 190;4 and 191;11 and 192;12 and 193;13 and 194;14 and 195;15 and 196;16 and 184;17 and 197;18 and 198;1 and 199;19 and 200 ;20 and 201;21 and 202;22 and 203;23 and 204;24 and 205;25 and 184;4 and 206;26 and 207;27 and 208;28 and 209;9 and 210;29 and 211;30 and 212;31 and 213;32 and 214;33 and 215;34 and 216;17 and 217;1 and 218;35 and 184;36 and 184;35 and 219;35 and 220;37 and 184;38 and 221;39 and 222;9 and 223;9 and 224;40 and 184;41 and 225;8 and 226;42 and 227;35 and 228;43 and 229;4 and 230;9 and 231;8 and 232;1 and 233;8 and 234;44 and 184;1 and 235;45 and 236;30 and 237;46 and 184;47 and 238;48 and 239;1 and 230;49 and 190;44 and 240;50 and 184;51 and 241;52 and 242;53 and 243;54 and 184;55 and 244;56 and 245;57 and 24 6;58 and 247;59 and 248;60 and 249;8 and 250;61 and 184;35 and 251;62 and 252;1 and 253;63 and 254;64 and 181;65 and 255;66 and 256;67 and 257;68 and 198;35 and 258;69 and 259;70 and 260;71 and 261;72 and 262;73 and 263;74 and 264;75 and 265;76 and 266;77 and 267;4 and 205;78 and 268;1 and 269;27 and 269;79 and 269; 80 and 270; 81 and 269; 22 and 269; 82 and 269; 83 and 271; 84 and 272; 1 and 273; 8 and 274; 8 and 275; 22 and 276; 8 and 277; 85 and 278; 86 and 279; 87 and 269; 88 and 280; 89 and 269; 60 and 281; 90 and 269; 91 and 282; 22 and 283; 92 and 269; 93 and 284; 94 and 285; 95 and 286; 96 and 269; 73 and 287; 97 and 288; 68 and 289; 98 and 269; 99 and 290; 100 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304; 72 and 305;114 and 306;115 and 307;116 and 269;117 and 308;118 and 269;110 and 309;119 and 310;120 and 311;121 and 269;122 and 312;123 and 289;9 and 313;124 and 314;120 and 289;125 and 315;110 and 316;126 and 317;127 and 318;49 and 319;128 and 320;9 and 321;22 and 322;129 and 323;130 and 269;131 and 324 ;132 and 325;133 and 326;134 and 327;53 and 328;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340;149 and 341; 150 and 342; 151 and 343; 152 and 344; 94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 3 Includes the VH and VL amino acid sequences described in 56;166 and 357;167 and 358;110 and 334;87 and 359;168 and 360;169 and 269;170 and 269;171 and 269;172 and 361;8 and 362;173 and 304;174 and 269;1 and 363;175 and 364;176 and 365;177 and 269;178 and 366;179 and 269;180 and 367; or 89 and 368.

[0093] In a particular embodiment, the amino acid sequences of VH and VL are, respectively, SEQ ID NOs: 1 and 181;2 and 182;3 and 183;4 and 184;5 and 185;1 and 184;6 and 184;1 and 186;7 and 187;8 and 188;9 and 189;10 and 190;4 and 191;11 and 192;12 and 193;13 and 194;14 and 195;15 and 196;16 and 184;17 and 197;18 and 198;1 and 199;19 and 200;20 and 201;21 and 202;2 2 and 203; 23 and 204; 24 and 205; 25 and 184; 4 and 206; 26 and 207; 27 and 208; 28 and 209; 9 and 210; 29 and 211; 30 and 212; 31 and 213; 32 and 214; 33 and 215; 34 and 216; 17 and 217; 1 and 218; 35 and 184; 36 and 184; 35 and 219; 35 and 220; 37 and 184; 38 and 221; 39 and 222; 9 and 223; 9 and 224; 40 and 184; 41 and 225; 8 and and 226;42 and 227;35 and 228;43 and 229;4 and 230;9 and 231;8 and 232;1 and 233;8 and 234;44 and 184;1 and 235;45 and 236;30 and 237;46 and 184;47 and 238;48 and 239;1 and 230;49 and 190;44 and 240;50 and 184;51 and 241;52 and 242;53 and 243;54 and 184;55 and 244;56 and 245;57 and 246;58 and 247;59 and 248 ;60 and 249;8 and 250;61 and 184;35 and 251;62 and 252;1 and 253;63 and 254;64 and 181;65 and 255;66 and 256;67 and 257;68 and 198;35 and 258;69 and 259;70 and 260;71 and 261;72 and 262;73 and 263;74 and 264;75 and 265;76 and 266;77 and 267;4 and 205;78 and 268;1 and 269;27 and 269;79 and 269;80 and 270;81 and 269; 22 and 269; 82 and 269; 83 and 271; 84 and 272; 1 and 273; 8 and 274; 8 and 275; 22 and 276; 8 and 277; 85 and 278; 86 and 279; 87 and 269; 88 and 280; 89 and 269; 60 and 281; 90 and 269; 91 and 282; 22 and 283; 92 and 269; 93 and 284; 94 and 285; 95 and 286; 96 and 269; 73 and 287; 97 and 288; 68 and 289; 98 and 269; 9 9 and 290; 100 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304; 72 and 305; 114 and 306; 115 and 307;116 and 269;117 and 308;118 and 269;110 and 309;119 and 310;120 and 311;121 and 269;122 and 312;123 and 289;9 and 313;124 and 314;120 and 289;125 and 315;110 and 316;126 and 317;127 and 318;49 and 319;128 and 320;9 and 321;22 and 322;129 and 323;130 and 269;131 and 324;132 and 325;133 and 326 ;134 and 327;53 and 328;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340;149 and 341;150 and 342;151 and 343; 152 and 344; 94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 356; 166 The amino acid sequences consist of those described as 357;167 and 358;110 and 334;87 and 359;168 and 360;169 and 269;170 and 269;171 and 269;172 and 361;8 and 362;173 and 304;174 and 269;1 and 363;175 and 364;176 and 365;177 and 269;178 and 366;179 and 269;180 and 367; or 89 and 368.

[0094] Any antibody format can be used in the antibodies disclosed herein. In certain embodiments, the antibody is a single-chain antibody or a single-chain Fv(scFv). In certain embodiments, the antibody is an scFv fused with an Fc region (scFv-Fc). In certain embodiments, the antibody is a Fab fragment. In certain embodiments, the antibody is an F(ab')2 fragment.

[0095] In certain embodiments, the antibodies or polypeptides disclosed herein are multispecific antibodies (e.g., bispecific antibodies) that specifically bind to both the PC and a second antigen.

[0096] In certain embodiments, the antibodies or polypeptides disclosed herein are conjugated to a cytotoxic agent, a cytostatic agent, a toxin, a radionuclide, or a detectable label. In certain embodiments, the cytotoxic agent is capable of inducing the death or destruction of cells with which it comes into contact. In certain embodiments, the cytostatic agent is capable of preventing or substantially reducing proliferation and / or inhibiting the activity or function of cells with which it comes into contact. In certain embodiments, the cytotoxic agent or cytostatic agent is a chemotherapeutic agent. In certain embodiments, the radionuclide is an isotope 3 H, 14 C, 32 P, 35 S, 36 Cl, 51 Cr, 57 Co, 58 Co, 59 Fe, 67 Cu, 90 Y, 99 Tc, 111 In, 117 Lu, 121 I, 124 I, 125 I, 131 I, 198 Au, 211 At, 213 Bi, 225 Ac, and 186 Re selected from the group consisting of. In certain embodiments, the detectable label comprises a fluorescent moiety or a click chemistry handle.

[0097] Any immunoglobulin (Ig) constant region can be used in the polypeptides and antibodies disclosed herein. In certain embodiments, the Ig region is a human IgG, IgE, IgM, IgD, IgA, or IgY immunoglobulin molecule, any class of immunoglobulin molecule (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG2a and IgG2b).

[0098] In certain embodiments, the Disclosure provides a polypeptide (e.g., an antibody) that specifically binds to a PC, the polypeptide comprising a heavy chain constant region optionally selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0099] In certain embodiments, the Disclosure provides a polypeptide (e.g., an antibody) that specifically binds to PC, wherein the polypeptide comprises a heavy chain constant region which is a variant of the wild-type heavy chain constant region, and the variant heavy chain constant region binds to FcγR with lower affinity than the wild-type heavy chain constant region binds to FcγR.

[0100] In certain embodiments, the disclosure provides an antibody that specifically binds to PC, the antibody comprising a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 683 or 684. In certain embodiments, the disclosure provides an antibody that specifically binds to PC, the antibody comprising a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 683 or 684. In certain embodiments, the disclosure provides an antibody that specifically binds to PC, the antibody comprising an allotype or isoallotype IgG1 heavy chain constant region selected from the group consisting of G1m1(a), G1m2(x), G1m3(f), and G1m17(z). See, for example, Jefferis and Lefranc (2009) mAbs 1(4):332-338.

[0101] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region (e.g., the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1, numbered according to the EU numbering system)) and / or the hinge region (residues 216-230, numbered according to the EU numbering system) of the antibodies described herein, thereby altering one or more functional properties of the antibody, such as serum half-life, complement binding, Fc receptor binding, and / or antigen-dependent cytotoxicity.

[0102] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced within the hinge region of the antibodies described herein, thereby altering (e.g., increasing or decreasing) the number of cysteine ​​residues in the hinge region, as described, for example, in U.S. Patent No. 5,677,425, which is incorporated herein by reference in its entirety. The number of cysteine ​​residues in the hinge region may be altered, for example, to facilitate the assembly of the light and heavy chains, or to alter (e.g., increase or decrease) the stability of the antibody.

[0103] In specific embodiments, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into the IgG constant region or its FcRn binding fragment (preferably an Fc or hinged Fc fragment) to alter (e.g., decrease or increase) the half-life of the antibody in vivo. For examples of mutations that alter (e.g., decrease or increase) the half-life of an antibody in vivo, see, for example, International Publication Nos. 02 / 060919, 98 / 23289, and 97 / 34631, and U.S. Patents Nos. 5,869,046, 6,121,022, 6,277,375, and 6,165,745 (all of which are incorporated herein by reference in their entirety). In certain embodiments, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into the IgG constant region or its FcRn binding fragment (preferably an Fc or hinged Fc fragment) to decrease the half-life of the antibody in vivo. In other embodiments, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into the IgG constant region or its FcRn binding fragment (preferably an Fc or hinged Fc fragment) to increase the half-life of the antibody in vivo. In specific embodiments, the antibody may have one or more amino acid mutations (e.g., substitutions) numbered according to the EU numbering system in a second constant (CH2) domain (residues 231-340 of human IgG1) and / or a third constant (CH3) domain (residues 341-447 of human IgG1). In specific embodiments, the constant region of IgG1 of the antibodies described herein includes substitutions numbered according to the EU numbering system: a substitution from methionine (M) to tyrosine (Y) at position 252, a substitution from serine (S) to threonine (T) at position 254, and a substitution from threonine (T) to glutamic acid (E) at position 256. See U.S. Patent No. 7,658,921, which is incorporated herein by reference in its entirety.This type of mutant IgG, referred to as the "YTE variant," has been shown to exhibit a four-fold increased half-life compared to the wild-type version of the same antibody (see Dall'Acqua WF et al., (2006) J Biol Chem 281:23514-24, which is incorporated herein by reference in its entirety). In certain embodiments, the antibody includes an IgG constant region containing one, two, three, or more amino acid substitutions of amino acid residues at positions 251–257, 285–290, 308–314, 385–389, and 428–436, numbered according to the EU numbering system.

[0104] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region (e.g., the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1, numbered according to the EU numbering system)) and / or the hinge region (residues 216-230, numbered according to the EU numbering system) of the antibodies described herein to increase or decrease the affinity of the antibody to the Fc receptor (e.g., an activated Fc receptor) on the surface of effector cells. Mutations in the Fc region of an antibody that decrease or increase the affinity of the antibody to the Fc receptor, and techniques for introducing such mutations into the Fc receptor or a fragment thereof, are known to those skilled in the art. Examples of mutations in the Fc receptor of antibodies that can be produced to alter the affinity of antibodies against the Fc receptor are described, for example, in Smith P et al., (2012) PNAS 109:6181-6186, U.S. Patent No. 6,737,056, and International Publication Nos. 02 / 060919, 98 / 23289, and 97 / 34631, all of which are incorporated herein by reference in their entirety.

[0105] In certain embodiments, the antibody comprises a heavy chain constant region that is a variant of the wild-type heavy chain constant region, the variant heavy chain constant region binding to FcγRIIB with higher affinity than the wild-type heavy chain constant region binds to FcγRIIB. In certain embodiments, the variant heavy chain constant region is a variant human heavy chain constant region, e.g., a variant human IgG1, a variant human IgG2, or a variant human IgG4 heavy chain constant region. In certain embodiments, the variant human IgG heavy chain constant region contains one or more of the following amino acid mutations according to the EU numbering system: G236D, P238D, S239D, S267E, L328F, and L328E. In certain embodiments, the variant human IgG heavy chain constant region includes a set of amino acid mutations selected from the group consisting of S267E and L328F, P238D and L328E, and P238D, according to the EU numbering system, as well as one or more substitutions selected from the group consisting of E233D, G237D, H268D, P271G, and A330R, P238D, E233D, G237D, H268D, P271G, and A330R, G236D and S267E, S239D and S267E, V262E, S267E, and L328F, and V264E, S267E, and L328F. In certain embodiments, FcγRIIB is expressed on cells selected from the group consisting of macrophages, monocytes, B cells, dendritic cells, endothelial cells, and activated T cells.

[0106] In further embodiments, one, two, or more amino acid substitutions are introduced into the IgG constant region Fc region to modify the effector function(s) of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 239, 243, 267, 292, 297, 300, 318, 320, 322, 328, 330, 332, and 396, numbered according to the EU numbering system, can be replaced with different amino acid residues so that the antibody has a modified affinity for the effector ligand, while the antigen-binding ability of the parent antibody is retained. The effector ligand whose affinity is altered can be, for example, the Fc receptor or the C1 component of complement. This approach is described in further detail in U.S. Patents 5,624,821 and 5,648,260, each of which is incorporated herein by reference in whole. In certain embodiments, deletion or inactivation of a constant region domain (by point mutation or other means) may reduce Fc receptor binding of the circulating antibody, thereby increasing tumor localization. For example, see U.S. Patents 5,585,097 and 8,591,886, each of which is incorporated herein by reference in its entirety. In certain embodiments, one or more amino acid substitutions are introduced into the Fc region of an antibody described herein to remove a potential glycosylation site on the Fc region, which may reduce Fc receptor binding (see, for example, Shields RL et al., (2001) J Biol Chem 276:6591-604, which is incorporated herein by reference in its entirety).In various embodiments, one or more of the following mutations in the constant region of the antibodies described herein may be produced: N297A substitution, N297Q substitution, L234A substitution, L234F substitution, L235A substitution, L235F substitution, L235V substitution, L237A substitution, S239D substitution, E233P substitution, L234V substitution, L235A substitution, C236 deletion, P238A substitution, S239D substitution, F243L substitution, D265A substitution, S267E substitution, L328F substitution, R292P substitution, Y300L substitution, A327Q substitution, P329A substitution, A330L substitution, I332E substitution, or P396L substitution, which are numbered according to the EU numbering system.

[0107] In certain embodiments, mutations selected from the group consisting of D265A, P329A, and combinations thereof, numbered according to the EU numbering system, may be produced in the constant region of the antibody described herein. In certain embodiments, mutations selected from the group consisting of L235A, L237A, and combinations thereof, numbered according to the EU numbering system, may be produced in the constant region of the antibody described herein. In certain embodiments, mutations selected from the group consisting of S267E, L328F, and combinations thereof, numbered according to the EU numbering system, may be produced in the constant region of the antibody described herein. In certain embodiments, mutations selected from the group consisting of S239D, I332E, optionally A330L, and combinations thereof, numbered according to the EU numbering system, may be produced in the constant region of the antibody described herein. In certain embodiments, mutations selected from the group consisting of L235V, F243L, R292P, Y300L, P396L, and combinations thereof, numbered according to the EU numbering system, may be produced in the constant region of the antibodies described herein. In certain embodiments, mutations selected from the group consisting of S267E, L328F, and combinations thereof, numbered according to the EU numbering system, may be produced in the constant region of the antibodies described herein.

[0108] In specific embodiments, the antibody described herein comprises a constant region of IgG1 having an N297Q or N297A amino acid substitution, numbered according to the EU numbering system. In particular embodiments, the antibody described herein comprises a constant region of IgG1 having a mutation selected from the group consisting of D265A, P329A, and combinations thereof, numbered according to the EU numbering system. In another embodiment, the antibody described herein comprises a constant region of IgG1 having a mutation selected from the group consisting of L234A, L235A, and combinations thereof, numbered according to the EU numbering system. In yet another embodiment, the antibody described herein comprises a constant region of IgG1 having a mutation selected from the group consisting of L234F, L235F, N297A, and combinations thereof, numbered according to the EU numbering system. In certain embodiments, the amino acid residues at positions L234, L235, and D265 of the human IgG1 heavy chain, numbered according to the EU numbering system, in the constant region of the antibody described herein are not L, L, and D, respectively. This approach is described in detail in International Publication No. 14 / 108483, which is incorporated herein by reference in its entirety. In certain embodiments, the amino acids corresponding to positions L234, L235, and D265 in the human IgG1 heavy chain are F, E, and A, or A, A, and A, respectively, and these are numbered according to the EU numbering system.

[0109] In certain embodiments, one or more amino acids selected from amino acid residues 329, 331, and 322 within the constant region of the antibody described herein, numbered according to the EU numbering system, can be replaced with different amino acid residues so that the antibody has modified C1q binding and / or reduced or absent complement-dependent cytotoxicity (CDC). This approach is described in more detail in U.S. Patent No. 6,194,551 (Idusogie et al.), which is incorporated herein by reference in its entirety. In certain embodiments, one or more amino acid residues within amino acid positions 231-238 of the N-terminal region of the CH2 domain of the antibody described herein, numbered according to the EU numbering system, are modified to alter the antibody's ability to immobilize complement. This approach is described in International Publication No. 94 / 29351 (which is incorporated herein by reference in its entirety). In certain embodiments, the Fc region of the antibody described herein is positioned at the following locations, 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, to increase the antibody's ability to mediate antibody-dependent cytotoxicity (ADCC) and / or to increase the antibody's affinity to the Fcγ receptor. These are modified by mutating one or more amino acids in 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 328, 329, 330, 331, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, ​​388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438, or 439 (e.g., by introducing amino acid substitutions), and these are numbered according to the EU numbering system. This approach is further described in International Publication No. 00 / 42072, which is incorporated herein by reference in its entirety.

[0110] In certain embodiments, the antibodies described herein include a modified constant region of IgG1, and the modification increases the antibody's ability to mediate antibody-dependent cytotoxicity (ADCC). In certain embodiments, as evaluated by the methods described herein and / or methods known to those skilled in the art, 0.1, 1, or 10 μg / mL of antibody is capable of inducing cell death of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% of PC-expressing cells within 1, 2, or 3 hours. In certain embodiments, the modified constant region of IgG1 includes S239D and I332E substitutions, numbered according to the EU numbering system. In certain embodiments, the modified constant region of IgG1 includes S239D, A330L, and I332E substitutions, numbered according to the EU numbering system. In certain embodiments, the modified constant region of IgG1 includes L235V, F243L, R292P, Y300L, and P396L substitutions, numbered according to the EU numbering system. In certain embodiments, the antibody is capable of inducing cell death in effector T cells and Tregs, where the percentage of Tregs undergoing cell death is at least 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4, 4.5, or 5 times higher than the percentage of effector T cells undergoing cell death.

[0111] In certain embodiments, the antibody described herein comprises a constant region of an IgG4 antibody, numbered according to the EU numbering system, in which serine at amino acid residue 228 of the heavy chain is substituted with proline. In certain embodiments, the disclosure provides an antibody that specifically binds to PC, the antibody comprising a heavy chain constant region having the amino acid sequence of SEQ ID NO: 684.

[0112] In certain embodiments, any of the constant region mutations or modifications described herein can be introduced into one or both heavy chain constant regions of an antibody described herein having two heavy chain constant regions.

[0113] Pharmaceutical composition Compositions comprising an anti-PC antibody disclosed herein having a desired purity in a physiologically acceptable carrier, excipient, or stabilizer are provided herein (see, for example, Remington's Pharmaceutical Sciences (1990), Mack Publishing Co., Easton, PA). Acceptable carriers, excipients, or stabilizers are nontoxic to the recipient at the dosage and concentration used and include buffers such as phosphates, citrates, and other organic acids, antioxidants including ascorbic acid and methionine, preservatives (e.g., octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl, or benzyl alcohol, alkylparabens such as methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol), low molecular weight (less than about 10 residues) polypeptides, serum albumin This includes proteins such as cellulose, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN®, PLURONIC®, or polyethylene glycol (PEG).

[0114] In specific embodiments, the pharmaceutical composition comprises an anti-PC antibody disclosed herein and optionally one or more additional prophylactic or therapeutic agents in a pharmaceutically acceptable carrier. In specific embodiments, the pharmaceutical composition comprises an anti-PC antibody disclosed herein and optionally one or more additional prophylactic or therapeutic agents in a pharmaceutically acceptable carrier. In certain embodiments, the antibody is the sole active ingredient in the pharmaceutical composition. The pharmaceutical compositions described herein may be useful for reducing or inhibiting PC activity and treating conditions such as inflammatory disorders. In certain embodiments, the disclosure relates to a pharmaceutical composition of the disclosure comprising the anti-PC antibody of the disclosure for use as a pharmaceutical. In other embodiments, the disclosure relates to a pharmaceutical composition of the disclosure for use in a method for treating inflammatory disorders.

[0115] Pharmaceutically acceptable carriers used in parenteral formulations include aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspensions and dispersants, emulsifiers, chelating agents, and other pharmaceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic glucose injection, sterile water injection, and glucose-added lactated Ringer's injection. Non-aqueous parenteral vehicles include plant-derived fixative oils, cottonseed oil, corn oil, sesame oil, and peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations, including phenol or cresol, mercury, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoates, thimerosal, benzalkonium chloride, and benzethonium chloride, can be added to parenteral formulations packaged in multi-dose containers. Examples of isotonic agents include sodium chloride and glucose. Examples of buffers include phosphates and citrates. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspensions and dispersants include sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. An emulsifier is polysorbate 80 (TWEEN® 80). A metal ion sequester or chelating agent is EDTA. The pharmaceutical carrier also includes ethyl alcohol, polyethylene glycol, and propylene glycol as water-miscible media, as well as sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0116] Pharmaceutical compositions can be formulated for any route of administration to a subject. Specific examples of routes of administration include intranasal, oral, pulmonary, transdermal, intradermal, and parenteral administration. Parenteral administration characterized by subcutaneous, intramuscular, or intravenous injection is also contemplated herein. Injectable preparations can be prepared in liquid solution or suspension, solid form suitable for dissolving or suspending in liquid before injection, or conventional forms such as emulsions. Injectable preparations, solutions, and emulsions also contain one or more excipients. Suitable excipients include, for example, water, saline, glucose, glycerol, or ethanol. In addition, if desired, the administered pharmaceutical composition may also contain wetting or emulsifying agents, pH buffers, stabilizers, solubility enhancers, and small amounts of non-toxic adjuvants such as other such agents, e.g., sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrin.

[0117] Formulations for parenteral antibody administration include sterile solutions ready for injection, sterile soluble products such as lyophilized powders ready for combination with a solvent immediately before use, sterile suspensions ready for injection, sterile insoluble products ready for combination with a vehicle immediately before use, and sterile emulsions. Solutions may be either aqueous or non-aqueous.

[0118] For intravenous administration, suitable carriers include physiological saline or phosphate-buffered saline (PBS), and solutions containing thickeners and solubilizers such as glucose, polyethylene glycol, and polypropylene glycol, as well as mixtures thereof.

[0119] Topical mixtures containing antibodies are prepared as described for topical and systemic administration. The resulting mixtures may be solutions, suspensions, emulsions, etc., and can be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, skin patches, or any other formulation suitable for topical administration.

[0120] The anti-PC antibodies disclosed herein can be formulated as aerosols for topical application, such as by inhalation (see, for example, U.S. Patents 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivering steroids useful in the treatment of inflammatory diseases, particularly asthma, and which are incorporated herein by reference in their entirety). These formulations for administration to the airways may be in the form of aerosols or solutions for nebulizers, or in the form of fine particle powders for inhalation, either alone or in combination with an inert carrier such as lactose. In such cases, the particles of the formulation will have a diameter of less than 50 microns in certain embodiments and less than 10 microns in certain embodiments.

[0121] The anti-PC antibodies disclosed herein can be formulated in the form of gels, creams, and lotions for topical or local application, for example, for topical application to mucous membranes such as the skin and inside the eyes, as well as for application to the eyes or intracapsular or intraspinal. Topical administration is considered for transdermal delivery, for administration to the eyes or mucous membranes, or for inhalation therapy. Intranasal solutions of the antibody alone or in combination with other pharmaceutically acceptable excipients can also be administered.

[0122] Transdermal patches, including iontophoresis and electrophoresis devices, are well known to those skilled in the art and can be used to administer antibodies. For example, such patches are disclosed in U.S. Patents 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010,715, 5,985,317, 5,983,134, 5,948,433, and 5,860,957, all of which are incorporated herein by reference.

[0123] In certain embodiments, the pharmaceutical compositions comprising the antibodies described herein are lyophilized powders that can be reconstituted for administration as solutions, emulsions, and other mixtures. They may also be reconstituted and formulated as solids or gels. The lyophilized powders are prepared by dissolving the antibodies described herein, or their pharmaceutically acceptable derivatives, in a suitable solvent. In certain embodiments, the lyophilized powders are sterile. The solvent may contain excipients or other pharmacological components that improve the stability of the powder or the reconstituted solutions prepared from the powders. Excipients that may be used include, but are not limited to, glucose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. The solvent may also contain buffers such as citrate, sodium phosphate, or potassium phosphate, or, in certain embodiments, other such buffers known to those skilled in the art at a nearly neutral pH. The desired formulations are obtained by subsequently aseptically filtering the solutions and then lyophilizing them under standard conditions known to those skilled in the art. In certain embodiments, the obtained solutions are divided into vials for lyophilization. Each vial contains a single-dose or multi-dose dose of the compound. Lyophilized powder can be stored under suitable conditions, such as from approximately 4°C to room temperature. Reconstitution of this lyophilized powder with sterile water for injection yields a formulation for parenteral administration. To reconstitute, the lyophilized powder is added to sterile water or another suitable carrier. The exact amount varies depending on the selected compound. Such amounts can be determined empirically.

[0124] The anti-PC antibodies disclosed herein and other compositions provided herein can also be formulated to target specific tissues, receptors, or other areas of the body being treated. Many of these targeting methods are well known to those skilled in the art. All such targeting methods are intended herein for use with the compositions of the present invention. Non-limiting examples of targeting methods include, for example, U.S. Patents No. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, and 6,0 See also Nos. 48,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874, all of which are incorporated herein by reference.

[0125] The composition used for in vivo administration can be sterile. This can be easily achieved, for example, by filtration through a sterile filtration membrane.

[0126] Instructions for use and usage In another aspect, the Disclosure provides a method for treating a subject using an anti-PC antibody disclosed herein. Any disease or disorder in a subject that would benefit from reduced oxidized phospholipid function can be treated using an anti-PC antibody disclosed herein.

[0127] In certain embodiments, the anti-PC antibodies disclosed herein are particularly useful for inhibiting the activity of PC in a subject. In certain embodiments, the anti-PC antibodies disclosed herein are particularly useful for inhibiting the inflammatory activity of PC in a subject. In certain embodiments, the anti-PC antibodies disclosed herein are particularly useful for treating inflammatory disorders or degenerative diseases. In certain embodiments, the anti-PC antibodies disclosed herein can be used as anti-inflammatory agents and / or anti-atherosclerotic agents. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat pain. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat osteoporosis. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat fibrotic diseases, including but not limited to pulmonary fibrosis. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat NETosis-driven diseases, including but not limited to anti-neutrophil cytoplasmic antibody (ANCA) vasculitis and type 1 diabetes mellitus. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat chronic obstructive pulmonary disease (COPD). In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat acute respiratory diseases, acute fulminant pneumonia, or other similar disorders. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat severe asthma. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat patients with acute hepatitis, or non-alcoholic steatohepatitis (NASH) and metabolic syndrome. In certain embodiments, the anti-PC antibodies disclosed herein can be used as anti-atherosclerotic agents for treating cardiovascular diseases and calcific aortic stenosis (CAS). In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat patients with acute coronary syndrome or patients with exacerbating angina.In certain embodiments, the anti-PC antibodies disclosed herein can be used as a preventive measure in the treatment of high-risk patients, such as patients with a stroke tendency or patients who develop atherosclerosis. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat high-risk patients with coronary artery disease (CAD). Accordingly, the anti-PC antibodies disclosed herein can be used to treat inflammatory diseases and disorders, cardiovascular diseases, and diseases related to oxidized phospholipids, including but not limited to atherosclerosis, acute coronary syndrome, acute myocardial infarction, myocardial infarction (heart attack), stable and unstable angina, aneurysms, coronary artery disease (CAD), ischemic heart disease, ischemic myocardium, cardiogenic death and sudden cardiac death, cardiomyopathy, congestive heart failure, heart failure, stenosis, peripheral artery disease (PAD), intermittent claudication, severe limb ischemia, and stroke. See U.S. Patent No. 11,008,381, which is incorporated herein by reference in its entirety.

[0128] In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat cardiovascular diseases, atherosclerosis, rheumatoid arthritis, osteoarthritis, lung tissue damage (e.g., caused by smoking), brain lesions, apoptosis, aging, Crohn's disease, ulcerative colitis, fatty liver disease (e.g., NASH), and non-alcoholic fatty liver disease (NAFLD). Therefore, the anti-PC antibodies disclosed herein can be used to treat inflammatory diseases and disorders, cardiovascular diseases, liver diseases and disorders (e.g., NASH, NAFLD), and diseases associated with oxidative stress and injury. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat non-alcoholic fatty liver disease. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat non-alcoholic steatohepatitis (NASH). In certain embodiments, the anti-PC antibodies disclosed herein can be used to inhibit the progression of NAFLD to NASH. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat or inhibit atherogenesis. See U.S. Patent No. 11,168,148, which is incorporated herein by reference in its entirety.

[0129] In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat reperfusion injury. In certain embodiments, reperfusion injury includes organ reperfusion injury. In certain embodiments, reperfusion injury includes ischemic reperfusion injury. In certain embodiments, organ reperfusion injury is induced by an ischemic event (e.g., myocardial infarction-induced reperfusion injury). In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat subjects at risk of or suffering from ischemic events, including, but not limited to, ischemic events associated with conditions selected from the group consisting of cerebral ischemia, intestinal ischemia, spinal cord ischemia, cardiovascular ischemia, myocardial ischemia associated with myocardial infarction, myocardial ischemia associated with congestive heart failure (CHF), ischemia associated with age-related macular degeneration (AME), hepatic ischemia, kidney / renal ischemia, skin ischemia, vasoconstrictive tissue ischemia, penile ischemia as a result of priapism and erectile dysfunction, ischemia associated with thromboembolic disease, ischemia associated with microvascular disease, ischemia associated with thrombosis, and ischemia associated with diabetic ulcers, gangrenous conditions, post-traumatic syndromes, cardiac arrest resuscitation, hypothermia, peripheral nerve injury, and neuropathy. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat subjects at risk of or already suffering from ischemic events resulting from induced injury, including but not limited to those from surgery, transplantation, accidental trauma, and mechanical support devices. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat subjects at risk of or already suffering from ischemic events resulting from cardiac surgery, kidney surgery, brain surgery, liver surgery, and bypass surgery. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat acute ischemic stroke. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat reperfusion-related indications, including organ transplantation such as liver, heart, and kidney transplantation.In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat thrombosis-related diseases, including but not limited to sickle cell disease, deep vein thrombosis, pulmonary embolism, cardiac embolism, hypercoagulation, thrombotic tendencies, factor V Leiden, antithrombin III deficiency, protein C deficiency, protein S deficiency, prothrombin gene mutation (G20210A), hyperhomocysteinemia, antiphospholipid syndrome (APS), anticardiolipin antibody (ACLA) thrombotic syndrome, and lupus anticoagulant (LA) syndrome. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat or inhibit atheromatosis. Please refer to U.S. Patent Application No. 16 / 759,331, which is incorporated herein by reference in its entirety.

[0130] In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat Toll-like receptor 2 (TLR2) mediated diseases or disorders, including but not limited to Kawasaki disease, including IVIG-refractory Kawasaki disease; type 2 diabetes; rheumatoid arthritis; skin diseases; multiple sclerosis; systemic lupus erythematosus; ulcerative colitis; Graves' disease; Sjögren's syndrome; autoimmune thyroid disease; or vasculitis. See U.S. Patent Application No. 16 / 965,271, which is incorporated herein by reference in its entirety.

[0131] In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat bacterial infections. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat neuroinflammatory diseases, including but not limited to amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), and Alzheimer's disease (AD). In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat Huntington's disease, frontotemporal dementia (FTD), multiple sclerosis (MS), clinical sporadic syndrome (CIS), and other demyelinating diseases of the central nervous system. In certain embodiments, the anti-PC antibodies disclosed herein can be used to treat eye diseases, including but not limited to age-related macular degeneration (AMD) and geographic atrophy (GA).

[0132] In certain embodiments, these methods further include administering an additional therapeutic agent to the subject. In certain embodiments, the additional therapeutic agent is an HMG-CoA reductase inhibitor (statin), an immunosuppressant, an ezetimide, a perixosome, nicotinic acid, a squalene inhibitor, a growth-activating receptor (PPAR) agonist, a cholesteryl ester transfer protein (CETP) inhibitor, an ApoA-1 Milano, a corticosteroid, a nonsteroidal anti-inflammatory drug, a steroidal anti-inflammatory drug, analgesics, a growth factor, an anti-atherosclerotic agent, an anti-proliferative agent, a HSP, a beta-2-glycoprotein-I, and any derivatives and analogues thereof.

[0133] HMGCoA reductase inhibitors (statins) are well-known drugs that effectively reduce low-density lipoprotein (LDL) cholesterol levels by inhibiting the enzyme that regulates the rate of cholesterol production, thereby increasing the clearance of LDL cholesterol (LDL-C) from the blood by the liver. Non-exclusive examples of commonly prescribed statins include atorvastatin, fluvastatin, lovastatin, pravastatin, and simvastatin.

[0134] Non-exclusive examples of immunosuppressants include biologics such as infliximab, abatacept, adalimumab, anakinra, certolizumab, etanercept, golimumab, ixekizumab, natalizumab, rituximab, secukinumab, tocilizumab, ustekinumab, canakinumab, and vedolizumab; calcineurin inhibitors such as cyclosporine and tacrolimus; mTOR inhibitors such as sirolimus and everolimus; inosine monophosphate dehydrogenase (IMDH) inhibitors such as azathioprine, leflunomide, and mycophenolate; monoclonal antibodies such as methotrexate, daclizumab, and basiliximab; muromonab-CD3; and Janus kinase inhibitors such as tofacitinib.

[0135] Ezetimibe is the first of a new class of cholesterol absorption inhibitors that potently and selectively inhibits dietary and bile cholesterol absorption at the brush border of the intestinal epithelium without affecting the absorption of triglycerides or fat-soluble vitamins. Thus, ezetimibe reduces overall cholesterol delivery to the liver, which in turn induces increased LDL receptor expression, leading to increased removal of LDL-C from the plasma.

[0136] Peroxisomes are single-membrane organelles present in almost all eukaryotic cells. One of the most important metabolic processes of peroxisomes is the β-oxidation of long-chain and very long-chain fatty acids. Peroxisomes are also involved in bile acid synthesis, cholesterol synthesis, plasmalogen synthesis, amino acid metabolism, and purine metabolism.

[0137] Nicotinic acid is a known drug that lowers total cholesterol, LDL cholesterol, and triglyceride levels while increasing high-density lipoprotein (HDL) cholesterol levels. There are three types of nicotinic acid drugs: immediate-release, time-release, and sustained-release. Nicotinic acid, or niacin, a water-soluble B vitamin, improves all lipoproteins when administered in doses far exceeding the vitamin requirement.

[0138] Squalene is an isoprenoid compound structurally similar to beta-carotene and is an intermediate metabolite in cholesterol synthesis. In humans, approximately 60% of dietary squalene is absorbed. It is transported in the serum in association with lipoproteins at very low densities and is ubiquitously distributed in human tissues, with the highest concentrations in the skin, which is one of the major components of skin surface lipids. Squalene inhibitors (e.g., monooxygenases and synthases) function as inhibitors of cholesterol biosynthesis.

[0139] Proliferation-activating receptor (PPAR) agonists, such as fibrates, are fatty acid-activating members of the nuclear receptor superfamily that play a crucial role in lipid and glucose metabolism and are involved in obesity-related metabolic diseases such as hyperlipidemia, insulin resistance, and coronary artery disease. Fibrates generally act as PPAR agonists, effectively reducing elevated plasma triglycerides and cholesterol. The most notable effect of fibrates includes a reduction in plasma triglyceride-rich lipoprotein (TRL). LDL-C levels generally decrease in individuals with elevated baseline plasma concentrations, while HDL cholesterol (HDL-C) levels typically increase when baseline plasma concentrations are low. Non-exclusive examples of commonly prescribed fibrates include bezafibrate, gemfibrozil, and fenofibrate.

[0140] Cholesteryl ester transfer protein (CETP) inhibitors play a major role in atherogenesis by reducing the accumulation of cholesteryl esters in macrophages and arterial walls, and therefore reduce foam cell formation, affecting cholesterol absorption. The most promising CETP inhibitor currently known is avisimibe.

[0141] ApoA-1 Milano is typically used as a recombinant complex with the phospholipid (ETC-216) to achieve significant regression of coronary artery atherosclerosis.

[0142] Non-steroidal anti-inflammatory drugs (NSAIDs) include, but are not limited to, oxicam, e.g., piroxicam, isoxicam, tenoxicam, sudoxicam, and CP-14,304; salicylates, e.g., aspirin, disalcid, benolilate, trilysate, sapphin, sorprin, diflunisal, and fendosal; acetic acid derivatives, e.g., diclofenac, fenclofenac, indomethacin, sulindac, tolmetin, isoxepac, flofenac, thiopinac, zidomethacin, acematacin, fenthiazac, zomepirac, clindanac, oxepinac, felbinac, and keto Lorac; Fenametes, e.g., mefenamic acid, meclofenamic acid, flufenamic acid, diflumic acid, and tolfenamic acid; Propionic acid derivatives, e.g., ibuprofen, naproxen, benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, thioxaprofen, suprofen, aluminoprofen, and thiaprofenic; Pyrazoles, e.g., phenylbutazone, oxyfenbutazone, feprazone, azapropazone, and trimethazone.

[0143] Non-exclusive examples of steroidal anti-inflammatory drugs include, but are not limited to, corticosteroids such as hydrocortisone, hydroxyltriamcinolone, alpha-methyldexamethasone, dexamethasone phosphate, beclomethasone dipropionate, clobetasol valerate, desonide, desoxymethasone, desoxycorticosterone acetate, dexamethasone, dichlorizone, diflorasone diacetate, diflucortolone valerate, fluadrenolone, fluchlorolone acetonide, fludrocortisone, flumetasone pivalate, fluocinolone acetonide, fluocinonide, flucortin butyl ester, fluocortolone, fluprednilidene acetate, flundrenolone, halcinonide, hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone, Examples include triamcinolone acetonide, cortisone, cortodoxone, flucetonide, fludrocortisone, difluorosone diacetate, fluradrenolon, fludrocortisone, difluorosone diacetate, fluradrenolon acetonide, medrizone, amsinafel, amsinafid, betamethasone and its esters in balance, chloroprednisone, chlorprednisone acetate, crocorterone, cresinolone, dichlorizone, diflurprednate, fluchloronide, flunisolide, fluoromethalone, fluperolon, fluprednisolone, hydrocortisone valerate, hydrocortisone cyclopentylpropionate, hydrocoltamate, meprednisone, paramethasone, prednisolone, prednisone, beclomethasone dipropionate, triamcinolone, and mixtures thereof.

[0144] Non-exclusive examples of analgesics (pain relievers) include aspirin and other salicylates (such as choline or magnesium salicylate), ibuprofen, ketoprofen, naproxen sodium, and acetaminophen.

[0145] Growth factors are hormones with numerous functions, including regulating adhesion molecule production, altering cell proliferation, increasing angiogenesis, enhancing collagen synthesis, regulating bone metabolism, and altering cell migration to a given region. Non-limiting examples of growth factors include insulin-like growth factor-1 (IGF-1), transforming growth factor-β (TGF-β), and bone morphogenetic proteins (BMPs).

[0146] Non-limiting examples of antiproliferative agents include alkylating agents, e.g., nitrogen mustard, ethyleneimine and methylmelamine, alkyl sulfonates, nitrosoureas, and triazenes; antimetabolites, e.g., folic acid analogs, pyrimidine analogs, and purine analogs; natural products, e.g., vinca alkaloids, epipodophyllotoxin, antibiotics, enzymes, taxanes, and biological response modifiers; various agents, e.g., platinum coordination complexes, anthracendions, anthracyclines, substituted ureas, methylhydrazine derivatives, or corticosteroids; or hormones or antagonists, e.g., corticosteroids, progestins, estrogens, antiestrogens, androgens, or gonadotropin-releasing hormone analogs. Specific examples of chemotherapeutic agents include nitrogen mustard, epipodophyllotoxin, antibiotics, platinum-coordinated complexes, bleomycin, doxorubicin, paclitaxel, etoposide, 4-OH cyclophosphamide, and cisplatinum.

[0147] The HSP family consists of approximately 25 proteins, distinguished by their molecular weight and highly conserved structures. Almost all humans have cellular and humoral immune responses to microbial heat shock protein 60 (HSP60). Because of the high antigenic homology between microorganisms (bacteria and parasites) and human HSP60, the "cost" of immunity to microorganisms may be the risk of cross-reactivity with human HSP60 expressed by stressed arterial endothelial cells. True autoimmunity to altered auto-HSP60 can also trigger this process (Wick et al., TRENDS in Immunology. 2001;22(12):665-669). HSPs are involved as target autoantigens in several experimental autoimmune diseases (arthritis, type 1 diabetes). Anti-HSP65 and anti-HSP60 antibodies are clearly associated with atherosclerotic lesions in humans. Studies conducted in rabbits and mice have shown that the generation of an HSP65-induced immune response by immunization with HSP65-rich preparations of recombinant protein or Mycobacterium tuberculosis enhances atherogenesis. As an autoimmune process pointing to HSP65 as a potential antigen candidate, a state of unresponsiveness is used to block these responses by inducing mucosal "tolerance." Therefore, our group reported that early atherosclerosis was attenuated in HSP65-ingested mice compared to either BSA-ingested or PBS-ingested mice (Harats et al., J Am Coll Cardiol. 2002;40:1333-1338), which was further supported by Maron, who demonstrated that nasal vaccination with HSP reduces inflammatory processes associated with atherosclerosis (Maron et al., Circulation. 2002;106:1708-1715).

[0148] Beta-2 glycoprotein I (beta-2GPI) is a phospholipid-binding protein that has been shown to function as a target for prethrombotic antiphospholipid antibodies. Recently, it has been demonstrated to drive immune-mediated responses and enhance mouse atherosclerosis. β-antibodies against beta-2-GPI have the ability to activate monocytes and endothelial cells, and can induce an immune response to beta-2GPI in accelerated atherosclerosis in mice susceptible to atherosclerosis. When beta-2GPI-responsive lymph nodes and spleen cells were transferred to LDL receptor-deficient mice, they promoted the formation of fatty streaks, demonstrating a direct pro-atherosclerotic role of beta-2GPI-specific lymphocytes. Inducing immune tolerance to beta-2GPI by prior oral administration of the antigen significantly reduced the degree of atherosclerotic lesions. Therefore, beta-2GPI is a candidate player in atherosclerotic plaques and may be usable as an immunomodulator of plaque progression. Oral administration of beta-2GPI inhibited the responsiveness of lymph node cells to beta-2GPI in mice immunized against human proteins. In lymph node cells of beta-2GPI-resistant mice immunized against beta-2GPI, priming with the respective proteins upregulated the production of IL-4 and IL-10. Therefore, oral administration of beta-2GPI is an effective means of suppressing atheroma formation in mice (George et al., Cardiovasc Res. 2004;62(3):603-9).

[0149] In certain embodiments, the anti-PC antibodies or pharmaceutical compositions disclosed herein may act synergistically with additional therapeutic agents.

[0150] In certain embodiments, the Disclosure relates to the Antibody and / or Pharmaceutical Composition of the Disclosure for use in the Method of the Disclosure, the Method further comprising administering an additional therapeutic agent to a subject. In certain embodiments, the Disclosure relates to (a) the Antibody and / or Pharmaceutical Composition of the Disclosure, and (b) an additional therapeutic agent for use as a pharmaceutical. In certain embodiments, the Disclosure relates to (a) the Antibody and / or Pharmaceutical Composition of the Disclosure, and (b) an additional therapeutic agent for use in a Method for the Treatment of Inflammatory Disorders. In further embodiments, the Disclosure relates to a pharmaceutical composition, kit or kit component comprising (a) the Antibody and / or Pharmaceutical Composition of the Disclosure, and (b) an additional therapeutic agent. In certain embodiments, additional therapeutic agents include HMG-CoA reductase inhibitors (statins), immunosuppressants, ezetimibe, peroxisomes, nicotinic acid, squalene inhibitors, growth-activating receptor (PPAR) agonists, cholesteryl ester transfer protein (CETP) inhibitors, ApoA-1 Milano, nonsteroidal anti-inflammatory drugs, steroidal anti-inflammatory drugs, analgesics, growth factors, anti-atherosclerotic agents, anti-proliferative agents, HSPs, or beta-2-glycoprotein-I.

[0151] Anti-PC antibodies and additional therapeutic agents (e.g., HMG-CoA reductase inhibitors (statins), immunosuppressants, ezetimibe, peroxisomes, nicotinic acid, squalene inhibitors, growth-activating receptor (PPAR) agonists, cholesteryl ester transfer protein (CETP) inhibitors, ApoA-1 Milano, nonsteroidal anti-inflammatory drugs, steroidal anti-inflammatory drugs, analgesics, growth factors, anti-atherosclerotic agents, antiproliferative agents, HSPs, or beta-2-glycoprotein-I) can be administered separately, sequentially, or simultaneously, in separate dosage forms.

[0152] The antibodies or pharmaceutical compositions described herein may be delivered to a target by various routes. These routes include, but are not limited to, parenteral, intranasal, intratracheal, oral, intradermal, topical, intramuscular, intraperitoneal, percutaneous, intravenous, intratumoral, conjunctival, intraarterial, and subcutaneous routes. Lung administration can also be used, for example, by the use of an inhaler or nebulizer, and by formulations using aerosolizing agents for use as a spray. In certain embodiments, the antibodies or pharmaceutical compositions described herein are delivered subcutaneously or intravenously. In certain embodiments, the antibodies or pharmaceutical compositions described herein are delivered intraarterially. In certain embodiments, the antibodies or pharmaceutical compositions described herein are delivered intratumorally. In certain embodiments, the antibodies or pharmaceutical compositions described herein are delivered to tumor inflow lymph nodes.

[0153] The amount of antibody or composition effective in treating and / or preventing a particular condition depends on the nature of the disease and can be determined by standard clinical techniques.

[0154] The precise dose used in a composition depends on the route of administration and the severity of the infection or disease it may cause, and should be determined according to the practitioner's judgment and the circumstances of each individual. For example, the effective dose may also vary depending on the means of administration, the target site, the patient's physiological state (including age, weight, and health), whether the patient is human or animal, other medications being administered, or whether the treatment is prophylactic or therapeutic. Typically, the patient is human, but non-human mammals, including transgenic mammals, can also be treated. The therapeutic dose may be titrated to optimize safety and efficacy.

[0155] The anti-PC antibodies described herein can also be used to assay OxPL levels in biological samples using classical immunohistochemical methods known to those skilled in the art, such as immunoassays including enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, or Western blotting. Suitable antibody assay labels are known in the art and include enzyme labels, e.g., glucose oxidase; radioisotopes, e.g., iodine. 125 I, 121 I), carbon ( 14 C), sulfur ( 35 S), tritium ( 3 H), Indium ( 121 In), and technetium ( 99 Examples of labels include luminescent labels such as Tc; luminescent labels such as luminol; and fluorescent labels such as fluorescein and rhodamine, and biotin. Such labels can be used to label the antibodies described herein. Alternatively, a second antibody that recognizes the anti-PC antibody described herein can be labeled and used in combination with the anti-PC antibody to detect OxPL levels. Thus, in certain embodiments, the disclosure relates to the use of the anti-PC antibody of the disclosure for in vitro detection of OxPL in a biological sample. In further embodiments, the disclosure relates to the use of the anti-PC antibody of the disclosure for assaying and / or detecting OxPL levels in a biological sample in vitro or ex vivo, wherein the anti-PC antibody is optionally conjugated to a radionuclide or a detectable label and / or carries a label described herein and / or an immunohistological method is used.

[0156] Assaying for OxPL levels is intended to involve measuring or estimating the level of OxPL in a first biological sample qualitatively or quantitatively, either directly (e.g., by determining or estimating an absolute OxPL level) or relatively (e.g., by comparing it to a disease-related OxPL level in a second biological sample). The OxPL level in the first biological sample can be measured or estimated and compared to a standard OxPL level, which is determined, for example, by taking a second biological sample from an individual without the disorder or by averaging the level from a population of individuals without the disorder. As is understood in the art, once a “standard” OxPL level is known, it can be repeatedly used as a standard for comparison. Therefore, in further embodiments, the present disclosure relates to in vitro methods for assaying and / or detecting OxPL levels in a biological sample, e.g., human OxPL levels, including measuring or estimating the level of OxPL in the biological sample qualitatively or quantitatively by immunohistological methods.

[0157] As used herein, the term “biological sample” refers to any biological sample obtained from an subject, cell line, tissue, or other cell source that may contain oxidized phospholipids. Methods for obtaining tissue biopsies and bodily fluids from animals (e.g., humans or cynomolgus monkeys) are well known in the art. Biological samples include peripheral blood mononuclear cells (PBMCs).

[0158] The anti-PC antibodies described herein are standard and well known to those skilled in the art and can be used for prognostic, diagnostic, monitoring, and screening applications, including in vitro and in vivo applications as described herein. Prognostic, diagnostic, monitoring, and screening assays and kits for in vitro assessment and measurement of immune system status and / or immune responses may be used to predict, diagnose, and monitor the evaluation of patient samples with respect to known or suspected immune system dysfunction, or to expected or desired immune system or antigen responses. Assessment and measurement of immune system status and / or immune responses can also be useful in determining whether a patient is suitable for clinical trials of a drug or for administration of a particular therapeutic agent or antibody (including combinations thereof) rather than different drugs or antibodies. In vivo applications include targeted cell therapy, and immune system modulation, and radioimaging of immune responses. Accordingly, in certain embodiments, this disclosure relates to the anti-PC antibodies and / or pharmaceutical compositions of this disclosure for use as diagnostic agents. In certain embodiments, the Disclosure relates to anti-PC antibodies and / or pharmaceutical compositions of the Disclosure for use in methods for predicting, diagnosing, and / or monitoring subjects having or suspected immune system dysfunction, and / or for predicting or desired immune system or antigenic responses. In other embodiments, the Disclosure relates to the use of anti-PC antibodies of the Disclosure for predicting, diagnosing, and / or monitoring subjects having or suspected immune system dysfunction, and / or for predicting or desired immune system or antigenic responses by in vitro or in vivo assays and / or detection of human OxPL levels in biological samples of subjects.

[0159] In certain embodiments, anti-PC antibodies can be used in immunohistochemistry of biopsy specimens. In certain embodiments, the method is an in vitro method. In another embodiment, anti-PC antibodies can be used to detect levels of OxPL, or levels of cells containing OxPL, which can then be associated with specific disease symptoms. The anti-PC antibodies described herein may carry detectable or functional labels and / or be conjugated to radionuclides or detectable labels. When fluorescent labels are used, specific binding members can be identified and quantified using currently available microscopy and fluorescence-activated cell sorter analysis (FACS), or a combination of both methods and procedures known in the art. The anti-PC antibodies described herein may carry or be conjugated to fluorescent labels. Exemplary fluorescent labels include, for example, reactive and conjugated probes, such as aminocoumarin, fluorescein, and Texas Red, Alexa Fluor dye, Cy dye, and DyLight dye. Anti-PC antibodies are isotopes 3 H, 14 C, 32 P, 35 S, 36 Cl, 51 Cr, 57 Co, 58 Co, 59 Fe, 67 Cu, 90 Y, 99 Tc, 111 In, 117 Lu, 121 I, 124 I, 125 I, 131 I, 198 Au, 211 At, 213 Bi, 225 Ac, and 186The anti-PC antibody may be radiolabeled or conjugated with radionuclides such as Re. When radiolabeling is used, the specific binding of the anti-PC antibody to PC can be identified and quantified using currently available counting procedures known in the art. If the label is an enzyme, detection can be achieved by any of the currently available colorimetric, spectrophotometric, fluorescence spectrophotometric, amperometric, or gas assay techniques known in the art. This can be achieved by contacting a sample or control sample with the anti-PC antibody under conditions that allow for the formation of a complex between the anti-PC antibody and PC. Any complex formed between the anti-PC antibody and PC is detected and compared between the sample and the control. PC can be specifically detected using the anti-PC antibody in light of the specific binding of the anti-PC antibodies described herein to PC. The anti-PC antibodies described herein can also be used to purify PC by immunoaffinity purification. Furthermore, this specification includes assay systems that can be prepared in the form of test kits, kits, or kit components for quantitatively analyzing the degree of PC / PC ligand complexes, for example. The system, test kit, kit, or kit component may include labeled components, such as labeled antibodies, and one or more additional immunochemical reagents.

[0160] Methods for producing polyparaploids, vectors, and antibodies In another embodiment, polynucleotides comprising nucleotide sequences encoding an antibody or a portion thereof as described herein, or fragments thereof that specifically bind to a PC antigen (e.g., VL and / or VH, and light and / or heavy chains), and vectors, for example, vectors comprising such polynucleotides for recombinant expression in host cells (e.g., E. coli and mammalian cells). Polynucleotides comprising nucleotide sequences encoding the heavy and / or light chains of any of the antibodies provided herein, and vectors comprising such polynucleotide sequences, for example, expression vectors for efficient expression in host cells, for example, mammalian cells, are provided herein.

[0161] As used herein, “isolated” polynucleotide or nucleic acid molecule is one isolated from other nucleic acid molecules present in a natural source of nucleic acid molecules (e.g., in mouse or human). Furthermore, “isolated” nucleic acid molecules, such as cDNA molecules, if produced by recombinant techniques, may substantially contain other cellular material or culture media, or if chemically synthesized, may substantially contain chemical precursors or other chemicals. For example, the language “substantially contained” includes preparations of polynucleotide or nucleic acid molecules containing less than about 15%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% (in particular less than about 10%) of other materials, such as cellular material, culture media, other nucleic acid molecules, chemical precursors, and / or other chemicals. In specific embodiments, the nucleic acid molecule(s) encoding the antibody described herein is isolated or purified.

[0162] In certain embodiments, polynucleotides are provided herein that include an antibody that specifically binds to a PC and comprises an amino acid sequence described herein, and a nucleotide sequence encoding an antibody that competes with such an antibody for binding to a PC (e.g., in a dose-dependent manner) or that binds to the same epitope as such an antibody.

[0163] In certain embodiments, this specification provides a polynucleotide comprising a nucleotide sequence encoding the light chain or heavy chain of an antibody described herein. The polynucleotide may include a nucleotide sequence encoding the light chain comprising the VL FR and CDR of an antibody described herein (see, for example, Table 1), or a nucleotide sequence encoding the heavy chain comprising the VH FR and CDR of an antibody described herein (see, for example, Table 1). In certain embodiments, the polynucleotide encodes the VH, VL, heavy chain, and / or light chain described herein. In another embodiment, the polynucleotide encodes the first VH and first VL described herein. In another embodiment, the polynucleotide encodes the second VH and second VL described herein. In another embodiment, the polynucleotide encodes the first heavy chain and first light chain described herein. In another embodiment, the polynucleotide encodes the second heavy chain and second light chain described herein. In another embodiment, the polynucleotide encodes the VH and / or VL, or the heavy chain and / or light chain of an antibody described herein.

[0164] Furthermore, this specification also provides polynucleotides encoding anti-PC antibodies optimized by, for example, codon / RNA optimization, substitution with heterologous signal sequences, and removal of mRNA instability elements. Methods for generating optimized nucleic acids encoding anti-PC antibodies or fragments thereof (e.g., light chain, heavy chain, VH domain, or VL domain) for recombinant expression by introducing codon changes and / or removing inhibitory regions in mRNA can be carried out, for example, by adapting the optimization methods described in U.S. Patents No. 5,965,726, No. 6,174,666, No. 6,291,664, No. 6,414,132, and No. 6,794,498 accordingly, all of which are incorporated herein by reference in their entirety. For example, potential splice sites and instability elements within RNA (e.g., A / T or A / U rich elements) can be mutated without altering the amino acids encoded by the nucleic acid sequence to increase the stability of the RNA for recombinant expression. The modification utilizes the degeneracy of the gene code, for example, by using alternative codons for the same amino acid. In certain embodiments, it may be desirable to alter one or more codons to encode conservative mutations, for example, similar amino acids that have a similar chemical structure and properties as well as / or function to the original amino acid. Such methods can increase the expression of anti-PC antibodies or fragments thereof by at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold or more compared to the expression of anti-PC antibodies encoded by unoptimized polynucleotides.

[0165] In certain embodiments, an optimized polynucleotide sequence encoding an anti-PC antibody or a fragment thereof (e.g., the VL domain and / or VH domain) described herein can hybridize to an antisense (e.g., complementary) polynucleotide of an unoptimized polynucleotide sequence encoding an anti-PC antibody or a fragment thereof (e.g., the VL domain and / or VH domain) described herein. In specific embodiments, an optimized nucleotide sequence encoding an anti-PC antibody or a fragment thereof described herein hybridizes to an antisense polynucleotide of an unoptimized polynucleotide sequence encoding an anti-PC antibody or a fragment thereof under high-stringency conditions. In specific embodiments, an optimized nucleotide sequence encoding an anti-PC antibody or a fragment thereof described herein hybridizes to an antisense polynucleotide of an unoptimized nucleotide sequence encoding an anti-PC antibody or a fragment thereof under high-stringency, medium-stringency, or low-stringency hybridization conditions. Information regarding hybridization conditions is provided, for example, in U.S. Patent Application Publication No. 2005 / 0048549 (e.g., paragraphs 72-73), which is incorporated herein by reference in its entirety.

[0166] Polynucleotides can be obtained by any method known in the art, and the nucleotide sequences of polynucleotides can be determined. The nucleotide sequences encoding the antibodies described herein, e.g., those listed in Table 1, and modified versions of these antibodies, can be determined using methods known in the art; that is, nucleotide codons known to encode specific amino acids are assembled in a manner that produces nucleic acids encoding antibodies. Polynucleotides encoding such antibodies can be assembled from chemically synthesized oligonucleotides (e.g., Kutmeier G et al., (1994), BioTechniques 17:242-6, which is incorporated herein by reference in its entirety), and in short, this involves the synthesis of duplicate oligonucleotides containing portions of the antibody-encoding sequence, annealing and ligation of these oligonucleotides, and then amplification of the ligated oligonucleotides by PCR.

[0167] Alternatively, the antigen-binding regions described herein or the polynucleotides encoding the antibodies described herein can be generated from nucleic acids from a suitable source (e.g., hybridomas) using methods well known in the art (e.g., PCR and other molecular cloning methods). For example, PCR amplification using synthetic primers hybridizable to the 3' and 5' ends of known sequences can be carried out using genomic DNA obtained from hybridoma cells producing the antibody of interest. Using such PCR amplification methods, nucleic acids containing sequences encoding the light and / or heavy chains of the antibody can be obtained. Using such PCR amplification methods, nucleic acids containing sequences encoding the variable light and / or variable heavy chain regions of the antibody can be obtained. The amplified nucleic acids can be cloned into vectors for expression in host cells and further cloning.

[0168] If a clone containing a nucleic acid encoding a specific antigen-binding region or antibody is unavailable, but the sequence of the antigen-binding region or antibody molecule is known, the nucleic acid encoding the immunoglobulin can be obtained, for example, from a cDNA library encoding an antibody by PCR amplification using synthetic primers that can hybridize to the 3' and 5' ends of the sequence, or by cloning using oligonucleotide probes specific to a particular gene sequence, either by chemical synthesis or from a suitable source (e.g., an antibody cDNA library, or a cDNA library produced from any tissue or cell expressing the antibody (such as hybridoma cells selected to express the antibody described herein), or nucleic acids isolated therefrom (preferably poly(A+RNA))). The cDNA clone can then be identified, for example, from an antibody-encoding cDNA library. The amplified nucleic acid produced by PCR can then be cloned into a replicable cloning vector using any method well known in the art.

[0169] The DNA encoding the anti-PC antibodies described herein can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to the genes encoding the heavy and light chains of the anti-PC antibody). Hybridoma cells can serve as a source of such DNA. Once isolated, the DNA is placed in an expression vector, which is then transfected into host cells such as E. coli cells, monkey COS cells, Chinese hamster ovary (CHO) cells (e.g., CHO cells from the CHO GS System® (Lonza)), or myeloma cells that do not otherwise produce immunoglobulin proteins, thereby obtaining the synthesis of anti-PC antibodies in recombinant host cells.

[0170] To generate a complete antibody or antigen-binding region, the VH or VL sequence within the scFv clone can be amplified using PCR primers containing the VH or VL nucleotide sequence, a restriction site, and an adjacent sequence to protect the restriction site. Using cloning techniques known to those skilled in the art, the PCR-amplified VH domain can be cloned into a vector expressing a heavy chain constant region, e.g., a human gamma 1 or human gamma 4 constant region, and the PCR-amplified VL domain can be cloned into a vector expressing a light chain constant region, e.g., a human kappa or lambda constant region. In certain embodiments, the vector for expressing the VH or VL domain includes an EF-1α promoter, a secretion signal, a cloning site for the variable region, a constant region, and a selection marker such as neomycin. The VH and VL domains can also be cloned into a single vector expressing the desired constant region. The heavy chain and light chain conversion vectors are then co-transfected into a cell line using techniques known to those skilled in the art to generate a stable or transient cell line expressing a full-length antibody, e.g., IgG.

[0171] DNA can also be modified, for example, by substituting the coding sequences of the human heavy and light chain constant regions for those of the mouse, or by covalently bonding all or part of the coding sequence of a non-immunoglobulin polypeptide to the coding sequence of an immunoglobulin.

[0172] Furthermore, polynucleotides that hybridize to the antibody-encoding polynucleotides described herein under high-stringency, medium-stringency, or low-stringency hybridization conditions are also provided. In specific embodiments, the polynucleotides described herein hybridize to the polynucleotides encoding the VH domain and / or VL domains provided herein under high-stringency, medium-stringency, or low-stringency hybridization conditions.

[0173] Hybridization conditions have been described in the art and are known to those skilled in the art. For example, hybridization under stringent conditions may include hybridization to filter-bound DNA in 6x sodium chloride / sodium citrate (SSC) at about 45°C, followed by one or more washes in 0.2x SSC / 0.1% SDS at about 50–65°C, and hybridization under highly stringent conditions may include hybridization to filter-bound nucleic acids in 6x SSC at about 45°C, followed by one or more washes in 0.1x SSC / 0.2% SDS at about 68°C. Hybridization under other stringent hybridization conditions is known and described to those skilled in the art; see, for example, Ausubel FM et al., eds., (1989) Current Protocols in Molecular Biology, Vol. I, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., New York, pp. 6.3.1–6.3.6 and 2.10.3, which are incorporated herein by reference in their entirety.

[0174] In certain embodiments, the Specified Information provides cells (e.g., host cells) that express (e.g., by recombination) the antibody described herein, which specifically binds to PC, along with associated polynucleotides and expression vectors. The Specified Information provides vectors (e.g., expression vectors) comprising polynucleotides containing nucleotide sequences encoding an anti-PC antibody or fragment for recombinant expression in host cells, preferably mammalian cells (e.g., CHO cells). The Specified Information also provides host cells containing such vectors for recombinant expression of the anti-PC antibody described herein (e.g., human or humanized antibody). In certain embodiments, the Specified Information provides methods for producing the antibody described herein, comprising expressing the antibody from host cells.

[0175] Recombinant expression of antibodies described herein that specifically bind to PC (e.g., the full-length antigen-binding region or antibody, or heavy and / or light chain, of an antibody described herein) typically involves the construction of an expression vector containing a polynucleotide encoding the antibody. Once a polynucleotide encoding an antibody molecule, an antibody heavy and / or light chain, or a fragment thereof (e.g., heavy and / or light chain variable region) is obtained, a vector for the production of the antibody molecule can be produced by recombinant DNA techniques using techniques well known in the art. Thus, methods for preparing proteins by expressing a polynucleotide containing an antibody or antibody fragment (e.g., light or heavy chain) encoding a nucleotide sequence are described herein. Using methods well known to those skilled in the art, an expression vector containing the antibody or antibody fragment (e.g., light or heavy chain) encoding sequence and appropriate transcription and translational regulatory signals can be constructed. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Also provided are replicable vectors containing nucleotide sequences that encode the antibody molecule, the heavy or light chain of the antibody, the heavy or light chain variable region of the antibody or a fragment thereof, or the CDR of the heavy or light chain, and are functionally linked to a promoter. Such vectors may, for example, contain a nucleotide sequence encoding the constant region of the antibody molecule (see, for example, International Publication Nos. 86 / 05807 and 89 / 01036, and U.S. Patent No. 5,122,464, all of which are incorporated herein by reference in their entirety), and the variable region of the antibody may be cloned into such a vector for the expression of the entire heavy chain, the entire light chain, or both the entire heavy chain and the entire light chain.

[0176] In certain embodiments, the vector comprises polynucleotides encoding the VH, VL, heavy chain, and / or light chain of the antibody described herein. In other embodiments, the vector comprises polynucleotides encoding the VH and VL of the antibody described herein. In other embodiments, the vector comprises polynucleotides encoding the heavy chain and light chain of the antibody described herein.

[0177] The expression vector can be transferred into cells (e.g., host cells) by conventional techniques, and the resulting cells can then be cultured by conventional techniques to produce antibodies or fragments thereof containing the antibodies or fragments thereof. Accordingly, the Specified provides host cells containing the antibodies or fragments thereof, their heavy or light chains, or fragments thereof, or polynucleotides encoding the single-chain antibodies thereof, functionally linked to a promoter for the expression of such sequences within the host cells.

[0178] In another embodiment, the host cell comprises a polynucleotide encoding VH and VL of the antibody described herein. In another embodiment, the host cell comprises a vector comprising a polynucleotide encoding VH and VL of the antibody described herein. In another embodiment, the host cell comprises a first polynucleotide encoding VH of the antibody described herein and a second polynucleotide encoding VL of the antibody described herein. In another embodiment, the host cell comprises a first vector comprising a first polynucleotide encoding VH of the antibody described herein and a second vector comprising a second polynucleotide encoding VL of the antibody described herein.

[0179] In specific embodiments, the heavy chain / heavy chain variable region expressed by a first cell associates with the light chain / light chain variable region of a second cell to form the anti-PC antibody described herein. In certain embodiments, a population of host cells including such a first host cell and such a second host cell is provided herein.

[0180] In certain embodiments, a vector population is provided herein, comprising a first vector containing a polynucleotide encoding the light chain / light chain variable region of an anti-PC antibody described herein, and a second vector containing a polynucleotide encoding the heavy chain / heavy chain variable region of an anti-PC antibody described herein.

[0181] Various host expression vector systems can be used to express the antibody molecules described herein (see, for example, U.S. Patent No. 5,807,715, which is incorporated herein by reference in its entirety). Such host expression systems represent a medium that can produce and subsequently purify the desired coding sequence, but also represent cells that, when transformed or transfected with the appropriate nucleotide coding sequence, can in situ express the antibody molecules described herein. These include microorganisms, such as bacteria (e.g., E. coli and B. subtilis), such as bacteria transformed with recombinant bacteriophage DNA, plasmid DNA, or cosmid DNA expression vectors containing antibody coding sequences; yeast (e.g., Saccharomyces and Pichia), such as yeast transformed with recombinant yeast expression vectors containing antibody coding sequences; insect cell lines, such as insect cell lines infected with recombinant virus expression vectors containing antibody coding sequences (e.g., baculovirus); plant cell lines (e.g., green algae, e.g., Chlamydomonas reinhardtii), such as plant cell lines infected with recombinant virus expression vectors containing antibody coding sequences (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV), or plant cell lines transformed with recombinant plasmid expression vectors containing antibody coding sequences (e.g., Ti plasmid); or mammalian cell lines (e.g., COS (e.g., COS1 or COS), CHO, BHK, MDCK, HEK) This includes, but is not limited to, mammalian cell lines possessing recombinant expression constructs containing promoters derived from mammalian cell genomes (e.g., metallothionein promoter) or promoters derived from mammalian viruses (e.g., late adenovirus promoter; vaccinia virus 7.5K promoter), such as 293, NS0, PER.C6, VERO, CRL7O3O, HsS78Bst, HeLa, and NIH 3T3, HEK-293T, HepG2, SP210, R1.1, BW, LM, BSC1, BSC40, YB / 20, and BMT10 cells), for example, mammalian cell lines possessing recombinant expression constructs containing promoters derived from mammalian cell genomes (e.g., metallothionein promoter) or mammalian viruses (e.g., late adenovirus promoter; vaccinia virus 7.5K promoter).In specific embodiments, the cells expressing the antibodies described herein are Chinese hamster ovary (CHO) cells, e.g., CHO cells from the CHO GS System (Lonza). In certain embodiments, the heavy and / or light chains of the antibodies produced by CHO cells may have an N-terminal glutamine or glutamate residue replaced by pyroglutamic acid. In certain embodiments, the cells expressing the antibodies described herein are human cells, e.g., human cell lines. In specific embodiments, the mammalian expression vector is pOptiVEC (trademark) or pcDNA3.3. In certain embodiments, bacterial cells such as Escherichia coli, or eukaryotic cells (e.g., mammalian cells), are used for the expression of recombinant antibody molecules, particularly for the expression of the whole recombinant antibody molecule. For example, mammalian cells, such as CHO cells, are a viable expression system for antibodies, along with vectors, such as the major early gene promoter element from human cytomegalovirus (Foecking MK & Hofstetter H (1986) Gene 45:101-5, and Cockett MI et al., (1990) Biotechnology 8(7):662-7; each of these publications is incorporated herein by reference in its entirety). In specific embodiments, the antibodies described herein are produced by CHO cells or NS0 cells. In specific embodiments, the expression of the nucleotide sequence encoding the antibodies described herein that specifically bind to PC is regulated by a constitutive promoter, an inducible promoter, or a tissue-specific promoter.

[0182] In bacterial systems, a number of expression vectors can be advantageously selected depending on the intended use of the expressed antibody molecule. For example, when producing large quantities of such antibodies for the creation of pharmaceutical compositions of antibody molecules, a vector that directs the expression of a high level of fusion protein product that can be easily purified may be desirable. Such vectors include, but are not limited to, the E. coli expression vector pUR278 (Ruether U & Mueller-Hill B (1983) EMBO J 2:1791-1794), in which the coding sequence can be individually ligated within the vector in a frame having a lac Z coding region, thereby producing a fusion protein; and pIN vectors (Inouye S & Inouye M (1985) Nuc Acids Res 13:3101-3109, Van Heeke G & Schuster SM (1989) J Biol Chem 24:5503-5509), and all of these literatures are incorporated herein by reference in their entirety. For example, pGEX vectors can also be used to express exogenous polypeptides as fusion proteins with glutathione 5-transferase (GST). Generally, such fusion proteins are soluble and can be readily purified from lysed cells by adsorption and binding to matrix glutathione agarose beads, followed by elution in the presence of free glutathione. pGEX vectors are designed to contain a thrombin or factor Xa protease cleavage site so that the cloned target gene product can be released from the GST portion.

[0183] In insect systems, for example, the Autographa californica nuclear polyhedron disease virus (AcNPV) can be used as a vector to express foreign genes. The virus proliferates in Spodoptera frugiperda cells. The coding sequence can be individually cloned into a non-essential region of the virus (e.g., the polyhedrin gene) and placed under the control of the AcNPV promoter (e.g., the polyhedrin promoter).

[0184] Many virus-based expression systems are available in mammalian host cells. When adenoviruses are used as expression vectors, the desired coding sequence can be ligated to the adenovirus transcription / translation regulatory complex, e.g., the late promoter and tripartite reader sequence. This chimeric gene can then be inserted into the adenovirus genome by in vitro or in vivo recombination. Insertion into non-essential regions of the viral genome (e.g., regions E1 or E3) results in a recombinant virus, which is viable and capable of expressing molecules in an infected host (see, e.g., Logan J & Shenk T (1984) PNAS 81(12):3655-9, which is incorporated herein by reference in its entirety). Efficient translation of the inserted coding sequence may also require specific start signals. These signals include the ATG start codon and adjacent sequences. Furthermore, to ensure translation of the entire insert, the start codon must be homeophase with the read frame of the desired coding sequence. These exogenous translational regulatory signals and start codons can have diverse origins, both natural and synthetic. Expression efficiency can be enhanced by including appropriate transcriptional enhancer elements, transcriptional terminators, etc. (see, for example, Bitter G et al., (1987) Methods Enzymol. 153:516-544, which is incorporated herein by reference in its entirety).

[0185] Furthermore, host cell lines can be selected that regulate the expression of the inserted sequence or modify and process the gene product in a desired, specific manner. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may be important for protein function. Different host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be selected to ensure that the precise modification and processing of expressed foreign proteins are achieved. For this purpose, eukaryotic host cells with cellular mechanisms for appropriate processing of primary transcripts, glycosylation, and phosphorylation of gene products can be used. Such mammalian host cells include, but are not limited to, CHO, VERO, BHK, Hela, MDCK, HEK 293, NIH 3T3, W138, BT483, Hs578T, HTB2, BT2O and T47D, NS0 (a mouse myeloma cell line that does not endogenously produce any immunoglobulin chains), CRL7O3O, COS (e.g., COS1 or COS), PER.C6, VERO, HsS78Bst, HEK-293T, HepG2, SP210, R1.1, BW, LM, BSC1, BSC40, YB / 20, BMT10, and HsS78Bst cells. In certain embodiments, the anti-PC antibodies described herein are produced in mammalian cells such as CHO cells.

[0186] In specific embodiments, the antibodies described herein have reduced or no fucose content. Such antibodies can be produced using techniques known to those skilled in the art. For example, the antibodies can be expressed in cells that lack or have no ability to fucosylate. In certain cases, antibodies with reduced fucose content can be produced using cell lines in which both alleles of α1,6-fucosyltransferase have been knocked out. The Potelligent® system (Lonza) is an example of such a system that can be used to produce antibodies with reduced fucose content.

[0187] To produce recombinant proteins in high yield over the long term, stable expression cells can be generated. For example, cell lines that stably express the anti-PC antibody described herein can be manipulated. In specific embodiments, the cells provided herein stably express light chain / light chain variable regions and heavy chain / heavy chain variable regions that associate to form the antigen-binding region or antibody described herein.

[0188] In certain embodiments, host cells can be transformed with DNA controlled by appropriate expression regulatory elements (e.g., promoters, enhancers, sequences, transcription terminators, polyadenylation sites, etc.) and selection markers, rather than using an expression vector containing a viral replication origin. After introducing the exogenous DNA / polynucleotide, the genetically engineered cells are grown in concentrated medium for 1-2 days, and then switched to selective medium. The selection markers in the recombinant plasmid confer resistance to selection, allowing the cells to stably incorporate the plasmid into their chromosomes, grow, and form lesions, which can then be cloned and grown into cell lines. This method can be advantageously used to manipulate cell lines expressing anti-PC or fragments thereof as described herein. Such engineered cell lines can be particularly useful for screening and measuring compositions that interact directly or indirectly with antibody molecules.

[0189] Many select systems can be used, including, but not limited to, the genes for herpes simplex virus thymidine kinase (Wigler M et al., (1977) Cell 11(1):223-32), hypoxanthine guanine phosphoribosyltransferase (Szybalska EH & Szybalski W (1962) PNAS 48(12):2026-2034), and adenine phosphoribosyltransferase (Lowy I et al., (1980) Cell 22(3):817-23) in tk-, hgprt-, or aprt- cells, and all of these are incorporated herein by reference in their entirety. Furthermore, antimetabolite resistance can be induced by the following genes: dhfr (Wigler M et al., (1980) PNAS 77(6):3567-70, O'Hare K et al., (1981) PNAS 78:1527-31) which induced resistance to methotrexate; gpt (Mulligan RC & Berg P (1981) PNAS 78(4):2072-6) which induced resistance to mycophenolate; and neo (Wu GY & Wu CH (1991) Biotherapy 3:87-95, Tolstoshev P (1993) Ann Rev Pharmacol Toxicol 32:573-596, Mulligan RC (1993) Science 260:926-932, and Morgan RA & Anderson) which induced resistance to aminoglycoside G-418. WF (1993) Ann Rev Biochem 62:191-217, Nabel GJ & Felgner PL (1993) Trends Biotechnol 11(5):211-5); and hygro (Santerre RF et al., (1984) Gene 30(1-3):147-56) which confers resistance to hygromycin can be used as a basis for selection, and all of these documents are incorporated herein by reference in their entirety.Methods commonly known in the field of recombinant DNA technology can be routinely applied to select desired recombinant clones, and such methods are described, for example, in Ausubel FM et al., (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993), Kriegler M, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990), and Chapters 12 and 13, Dracopoli NC et al., (eds.), Current Protocols in Human Genetics, John Wiley & Sons, NY (1994), Colbere-Garapin F et al., (1981) J Mol Biol 150:1-14, all of which are incorporated herein by reference in their entirety.

[0190] The expression level of antibody molecules can be increased by vector amplification (see Bebbington CR & Hentschel CCG, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol. 3 (Academic Press, New York, 1987), which is incorporated herein by reference in its entirety). If the marker in the vector system is amplified, an increase in the level of the inhibitor present in the host cell culture increases the copy number of the marker gene. Furthermore, since the amplified region is related to the target gene, protein production increases (see Crouse GF et al., (1983) Mol Cell Biol 3:257-66, which is incorporated herein by reference in its entirety).

[0191] Host cells can be co-transfected with two or more expression vectors described herein: a first vector encoding a heavy-chain polypeptide and a second vector encoding a light-chain polypeptide. The two vectors may contain identical selection markers that enable equivalent expression of the heavy-chain and light-chain polypeptides. Host cells can be co-transfected with two or more expression vectors in different amounts. For example, host cells can be transfected with any of the following ratios of the first expression vector to the second expression vector: approximately 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, or 1:50.

[0192] Alternatively, it is possible to use a single vector capable of encoding and expressing both heavy and light chain polypeptides. In such situations, the light chain should be placed before the heavy chain to avoid an excess of non-toxic free heavy chains (Proudfoot NJ (1986) Nature 322:562-565, and Koehler G (1980) PNAS 77:2197-2199, each of these publications is incorporated herein by reference in its entirety). The coding sequences for the heavy and light chains can include cDNA or genomic DNA. Expression vectors can be monocistronic or multicistronic. Multicistronic nucleic acid constructs can encode 2, 3, 4, 5, 6, 7, 8, 9, 10 or more gene / nucleotide sequences, or in the range of 2-5, 5-10, or 10-20 gene / nucleotide sequences. For example, a bisistronic nucleic acid construct may include, in the following order: a promoter, a first gene (e.g., the heavy chain of an antibody described herein), and a second gene (e.g., the light chain of an antibody described herein). In such an expression vector, the transcription of both genes may be driven by the promoter, the translation of mRNA from the first gene may be by a cap-dependent scanning mechanism, and the translation of mRNA from the second gene may be by a cap-independent mechanism, such as IRES.

[0193] Once the antibody molecules described herein are produced by recombinant expression, they can be purified by any method known in the art for the purification of immunoglobulin molecules, for example, by chromatography (e.g., by ion exchange, affinity, particularly affinity for specific antigens after protein A, and by sizing column chromatography), centrifugation, differential solubility, or any other standard technique for protein purification. Furthermore, to facilitate purification, the antibodies described herein can be fused to heterologous polypeptide sequences described herein or known in the art.

[0194] In specific embodiments, the antibodies described herein are isolated or purified. In certain embodiments, the isolated antibody is an antibody that substantially does not contain other antibodies having different antigen specificity from the isolated antibody. For example, in certain embodiments, the antibody preparations described herein are substantially free of cellular material and / or chemical precursors. The language “substantially free of cellular material” includes antibody preparations in which the antibody is isolated or separated from cellular components of cells produced by recombination. Thus, a substantially cellular material-free antibody includes antibody preparations having less than 30%, 20%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% (by dry weight) of heterologous proteins (also referred herein as “contamination proteins”) and / or variants of the antibody, e.g., different post-translational modified forms of the antibody, or other different forms of the antibody (e.g., antibody fragments). When antibodies are produced recombinantly, they generally contain substantially no culture medium; that is, the culture medium accounts for less than about 20%, 10%, 2%, 1%, 0.5%, or 0.1% of the volume of the protein preparation. When antibodies are produced by chemosynthesis, they generally contain substantially no chemical precursors or other chemicals; that is, they are separated from chemical precursors or other chemicals involved in protein synthesis. Thus, such preparations of antibodies contain less than about 30%, 20%, 10%, or 5% (by dry weight) of chemical precursors or compounds other than the antibody of interest. In specific embodiments, the antibodies described herein are isolated or purified.

[0195] Anti-PC antibodies or fragments thereof can be produced by any method known in the art for the synthesis of proteins or antibodies, such as chemical synthesis or recombinant expression techniques. Unless otherwise indicated, the methods described herein utilize conventional techniques in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and related fields within the art. These techniques are described and fully explained in the references cited herein, for example.For example, Maniatis T et al., (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Sambrook J et al., (1989), Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, Ausubel FM et al. al.,Current Protocols in Molecular Biology,John Wiley&Sons(1987 and annual updates),Current Protocols in Immunology,John Wiley&Sons(1987 and annual updates)Gait(ed.)(1984)Oligonucleotide Synthesis:A Practical Approach,IRL Press;Eckstein(ed.)(1991)Oligonucleotides and Analogues:A Practical Approach,IRL Press, Birren B et See al., (eds.) (1999) Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press. All of these documents are incorporated herein by reference in their entirety.

[0196] In specific embodiments, the antibodies described herein are antibodies prepared, expressed, produced, or isolated by any means, such as synthesis, or by genetic manipulation of DNA sequences. In certain embodiments, such antibodies contain sequences (e.g., DNA sequences or amino acid sequences) that are not naturally present in the in vivo antibody germline repertoire of animals or mammals (e.g., humans).

[0197] In one embodiment, a method for producing an anti-PC antibody is provided herein, comprising culturing cells or host cells described herein. In certain embodiments, the method is carried out in vitro. In certain embodiments, a method for producing an anti-PC antibody is provided herein, comprising expressing (e.g., recombinantly expressing) the antibody using cells or host cells described herein (e.g., cells or host cells containing polynucleotides encoding the antibody described herein). In certain embodiments, the cells are isolated cells. In certain embodiments, the exogenous polynucleotides are introduced into the cells. In certain embodiments, the method further comprises the step of purifying the antibody obtained from the cells or host cells.

[0198] In certain embodiments, the antibody is produced by expressing the polynucleotides encoding the VH and VL of the antibody described herein in a cell under conditions suitable for polynucleotide expression and antibody production. In other embodiments, the antibody is produced by expressing the polynucleotides encoding the heavy chain and light chain of the antibody described herein in a cell under conditions suitable for polynucleotide expression and antibody production. In certain embodiments, the antibody is produced by expressing a first polynucleotide encoding the VH of the antibody described herein and a second polynucleotide encoding the VL of the antibody described herein in a cell under conditions suitable for polynucleotide expression and antibody production. In certain embodiments, the antibody is produced by expressing a first polynucleotide encoding the heavy chain of the antibody described herein and a second polynucleotide encoding the light chain of the antibody described herein in a cell under conditions suitable for polynucleotide expression and antibody production.

[0199] Methods for producing polyclonal antibodies are known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel FM et al., eds., John Wiley and Sons, New York, which is incorporated herein by reference in its entirety).

[0200] Monoclonal antibodies can be prepared using a wide range of techniques known in the art, including the use of hybridoma, recombination, and phage display techniques, or combinations thereof. For example, monoclonal antibodies can be produced using hybridoma techniques, including techniques known in the art, which are taught, for example, in Harlow E & Lane D, Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed. 1988) and Hammerling GJ et al., Monoclonal Antibodies and T-Cell Hybridomas 563 681 (Elsevier, NY, 1981), each of which is incorporated herein by reference in whole. As used herein, the term “monoclonal antibody” is not limited to antibodies produced by hybridoma techniques. For example, monoclonal antibodies can be recombinantly produced from antibodies or fragments thereof described herein, for example, from host cells exogenously expressing the light and / or heavy chains of such antibodies.

[0201] In specific embodiments, as used herein, “monoclonal antibody” is an antibody produced by a single cell (e.g., a hybridoma or host cell that produces recombinant antibodies), and the antibody is specifically bound to a phage cell (PC) as determined, for example, by ELISA or other antigen-binding assays known in the art or shown in the examples provided herein. In certain embodiments, the monoclonal antibody may be a chimeric antibody or a humanized antibody. In certain embodiments, the monoclonal antibody may be a monovalent or polyvalent (e.g., bivalent) antibody. In certain embodiments, the monoclonal antibody may be a monospecific antibody or a multispecific antibody (e.g., bispecific antibody). The monoclonal antibodies described herein may be prepared, for example, by the hybridoma method described in Koehler G & Milstein C (1975) Nature 256:495 (which is incorporated herein by reference in its entirety), or they may be isolated, for example, from a phage library using the techniques described herein. Other methods for preparing clonal cell lines and the monoclonal antibodies expressed thereby are well known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel FM et al., cited above).

[0202] As used herein, an antibody is polyvalent (e.g., bivalent) bound to an antigen if it comprises at least two (e.g., two or more) monovalent binding regions, and each monovalent binding region is capable of binding to an epitope on the antigen. Each monovalent binding region can bind to the same or different epitopes on the antigen.

[0203] Methods for producing and screening specific antibodies using hybridoma technology are commonplace and well known in the art. For example, in the hybridoma method, mice, or other suitable host animals such as sheep, goats, rabbits, rats, hamsters, and macaques, are immunized to induce lymphocytes that produce or are capable of producing antibodies that specifically bind to the protein used for immunization (e.g., PC). Alternatively, lymphocytes may be immunized in vitro. The lymphocytes are then fused with myeloma cells using a suitable fusion agent such as polyethylene glycol to form hybridoma cells (Goding JW (ed.), Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986) (the entire text is incorporated herein by reference)). Furthermore, animals can be immunized using the RIMMS (Repeated Immunization Multiple Site) technique (Kilpatrick KE et al., (1997) Hybridoma 16:381-9 (the entire article is incorporated herein by reference)).

[0204] In certain embodiments, mice (or other animals such as rats, monkeys, donkeys, pigs, sheep, hamsters, or dogs) can be immunized with an antigen (e.g., PC), and once an immune response is detected, for example, when antigen-specific antibodies are detected in the mouse serum, the mouse spleen is collected and splenocytes are isolated. The splenocytes are then fused with any suitable myeloma cells, for example, cells from the SP20 cell line available from the American Type Culture Collection (ATCC®) (Manassas, VA), by known techniques to form hybridomas. The hybridomas are selected and cloned by limiting dilution. In certain embodiments, lymph nodes from immunized mice are collected and fused with NS0 myeloma cells.

[0205] The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the proliferation or survival of unfused parent myeloma cells. For example, if parent myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT), the hybridoma culture medium typically contains hypoxanthine, aminopterin, and thymidine (HAT medium), which inhibit the proliferation of HGPRT-deficient cells.

[0206] Specific embodiments utilize myeloma cells that efficiently fuse, support stable high-level antibody production by selected antibody-producing cells, and are sensitive to culture media such as HAT medium. These myeloma cell lines include NS0 cell lines and myeloma cell lines available from the Salk Institute Cell Distribution Center in San Diego, California, USA, such as MOPC-21 and MPC-11 mouse tumor-derived cell lines, as well as SP-2 or X63-Ag8.653 cells available from the American Type Culture Collection in Rockville, Maryland, USA. Human myeloma and mouse-human heterozygous myeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor D (1984) J Immunol 133:3001-5, Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987) (each of these is incorporated herein by reference in whole)).

[0207] The culture medium in which hybridoma cells are proliferating is assayed for the production of monoclonal antibodies against PC. The binding specificity of the monoclonal antibodies produced by hybridoma cells is determined by methods known in the art, such as immunoprecipitation, or in vitro binding assays such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA).

[0208] After hybridoma cells producing antibodies with desired specificity, affinity, and / or activity are identified, clones can be subcloned by limiting dilution procedures and proliferated by standard methods (Goding JW (ed.), Monoclonal Antibodies: Principles and Practice, cited above). Suitable culture media for this purpose include, for example, D-MEM or RPMI1640 medium. Furthermore, hybridoma cells can be proliferated in vivo as ascites tumors in animals.

[0209] Monoclonal antibodies secreted by subclones can be suitably separated from culture media, ascites fluid, or serum by conventional immunoglobulin purification procedures such as protein A-Sepharose chromatography, hydroxyl apatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0210] The antibodies described herein include, for example, antibody fragments that recognize PC, which can be produced by any technique known to those skilled in the art. For example, the Fab and F(ab')2 fragments described herein can be produced by proteolytic cleavage of immunoglobulin molecules using enzymes such as papain (for producing the Fab fragment) or pepsin (for producing the F(ab')2 fragment). The Fab fragment corresponds to one of the two identical arms of the antibody molecule and contains a complete light chain paired with the VH and CH1 domains of the heavy chain. The F(ab')2 fragment contains two antigen-binding arms of the antibody molecule linked by a disulfide bond in the hinge region.

[0211] Furthermore, the antibodies described herein can also be generated using various phage display methods known in the art. In phage display methods, a functional antibody domain is displayed on the surface of a phage particle carrying the polynucleotide sequence encoding it. In particular, the DNA sequences encoding the VH and VL domains are amplified from an animal cDNA library (e.g., a human or mouse cDNA library of diseased tissue). The DNA encoding the VH and VL domains is recombinant with an scFv linker by PCR and cloned into a phagemide vector. The vector is electroporated into E. coli to infect E. coli with helper phages. The phages used in these methods are typically filamentous phages containing fd and M13, and the VH and VL domains are usually recombinantly fused to either phage gene III or gene VIII. Phages expressing antigen-binding regions that bind to specific antigens can be selected or identified by antigen, for example, using a labeled antigen or an antigen bound to or captured on a solid surface or beads.Examples of phage display methods that can be used to produce the antibodies described herein are disclosed below: Brinkman U et al., (1995) J Immunol Methods 182:41-50, Ames RS et al., (1995) J Immunol Methods 184:177-186, Kettleborough CA et al., (1994) Eur J Immunol 24:952-958, Persic L et al., (1997) Gene 187:9-18, Burton DR & Barbas CF (1994) Advan Immunol 57:191-280, PCT applications PCT / GB91 / 001134, International Publications 90 / 02809, 91 / 10737, 92 / 01047, 92 / 18619, 93 / 11236, 95 / 15982, 95 / 20401, and 97 / 13844, and U.S. Patents 5,698,426, 5,223,409, and 5,403,484 , Nos. 5,580,717, 5,427,908, 5,750,753, 5,821,047, 5,571,698, 5,427,908, 5,516,637, 5,780,225, 5,658,727, 5,733,743, and 5,969,108 (all of these are incorporated herein by reference in their entirety).

[0212] As described in the above references, after phage selection, the antibody-coding region from the phage is isolated and used to generate the whole antibody, including human antibodies, or other desired antigen-binding fragments, which can then be expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, as described below. Techniques for recombinant production of antibody fragments such as Fab, Fab', and F(ab')2 fragments can also be used using methods known in the art, such as those disclosed in PCT Publication No. 92 / 22324, Mullinax RL et al., (1992) BioTechniques 12(6):864-9, Sawai H et al., (1995) Am J Reprod Immunol 34:26-34, and Better M et al., (1988) Science 240:1041-1043, all of which are incorporated herein by reference in their entirety.

[0213] In certain embodiments, the VH or VL sequence can be amplified from a template, e.g., an scFv clone, using PCR primers containing the VH or VL nucleotide sequence, a restriction site, and an adjacent sequence to protect the restriction site, in order to generate a complete antibody. Using cloning techniques known to those skilled in the art, the PCR-amplified VH domain can be cloned into a vector expressing the VH constant region, and the PCR-amplified VL domain can be cloned into a vector expressing the VL constant region, e.g., the human kappa or lambda constant region. The human kappa and lambda constant region sequences are known to those skilled in the art. Examples of human kappa and lambda constant region sequences are shown in SEQ ID NOs. 688-693. The VH and VL domains can also be cloned into a single vector expressing the desired constant region. The heavy-chain and light-chain conversion vectors are then co-transfected into a cell line using techniques known to those skilled in the art to generate a stable or transient cell line expressing a full-length antibody, e.g., IgG.

[0214] A chimeric antibody is a molecule in which different parts of the antibody are derived from different immunoglobulin molecules. For example, a chimeric antibody may include a variable region of a mouse or rat monoclonal antibody fused to the constant region of a human antibody. Methods for producing chimeric antibodies are known in the art. See, for example, Morrison SL (1985) Science 229:1202-7, Oi VT & Morrison SL (1986) BioTechniques 4:214-221, Gillies SD et al., (1989) J Immunol Methods 125:191-202, and U.S. Patents No. 5,807,715, No. 4,816,567, No. 4,816,397, and No. 6,331,415, all of which are incorporated herein by reference in their entirety.

[0215] A humanized antibody is capable of binding to a given antigen and comprises a framework region having substantially the amino acid sequence of a human immunoglobulin and a CDR having substantially the amino acid sequence of a non-human immunoglobulin (e.g., mouse immunoglobulin). In certain embodiments, the humanized antibody also includes an immunoglobulin constant region (Fc), typically at least a portion of the constant region (Fc) of a human immunoglobulin. The antibody may also include the CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain. The humanized antibody can be selected from any class of immunoglobulin, including IgM, IgG, IgD, IgA, and IgE, and any isotype, including IgG1, IgG2, IgG3, and IgG4.Humanized antibodies are used in CDR grafts (European Patent No. 239400, International Publication No. 91 / 09967, and U.S. Patents No. 5,225,539, 5,530,101, and 5,585,089), veneering, or resurfacing (European Patents No. 592106 and 519596, Padlan EA (1991) Mol Immunol 28(4 / 5):489-498, Studnicka GM et al., (1994) Prot Engineering 7(6):805-814, and Roguska MA et al., (1994) PNAS 91:969-973), chain shuffling (U.S. Pat. No. 5,565,332), and, for example, U.S. Pat. al.,(2000)Protein Eng.13(5):353-60, Morea V et al.,(2000)Methods 20(3):267-79, Baca M et al.,(1997)J Biol Chem 272(16):10678-84, Roguska MA et al.,(1996)Protein Eng 9(10):895 904, Couto JR et. The materials can be produced using a variety of known techniques, including, but not limited to, those disclosed in al., (1995) Cancer Res. 55 (23 Supp): 5973-5977s, Couto JR et al., (1995) Cancer Res 55(8): 1717-22, Sandhu JS (1994) Gene 150(2): 409-10, and Pedersen JT et al., (1994) J Mol Biol 235(3): 959-73, all of which are incorporated herein by reference in their entirety. See also U.S. Patent Publication No. 2005 / 0042664 A1 (February 24, 2005) and International Publication No. 2008 / 020079, both of which are incorporated herein by reference in their entirety.

[0216] Methods for producing multispecific antibodies (e.g., bispecific antibodies) are described, and refer to, for example, U.S. Patents 7,951,917, 7,183,076, 8,227,577, 5,837,242, 5,989,830, 5,869,620, 6,132,992, and 8,586,713, all of which are incorporated herein by reference in their entirety.

[0217] Bispecific, bivalent antibodies and methods for producing them are described, for example, in U.S. Patent Nos. 5,731,168, 5,807,706, 5,821,333, and U.S. Patent Publication Nos. 2003 / 020734 and 2002 / 0155537, each of which is incorporated herein by reference in its entirety. Bispecific, tetravalent antibodies and methods for producing them are described, for example, in International Patent Publication Nos. 02 / 096948 and 00 / 44788, both of which are incorporated herein by reference in their entirety. Generally, please refer to International Patent Application Publications 93 / 17715, 92 / 08802, 91 / 00360, and 92 / 05793, Tutt et al., J.Immunol. 147:60-69 (1991), U.S. Patents 4,474,893, 4,714,681, 4,925,648, 5,573,920, and 5,601,819, and Kostelny et al., J.Immunol. 148:1547-1553 (1992), each of which is incorporated herein by reference in its entirety.

[0218] The bispecific antibodies described herein can be produced, for example, according to the DuoBody technology platform (Genmab A / S) described in International Publication Nos. 2011 / 131746, 2011 / 147986, 2008 / 119353, and 2013 / 060867, and Labrijn AF et al., (2013) PNAS 110(13):5145-5150. Using DuoBody technology, a first monospecific antibody containing two heavy chains and two light chains, or half of the first antigen-binding region, can be combined with a second monospecific antibody containing two heavy chains and two light chains, or half of the second antigen-binding region. The resulting heterodimer contains one heavy chain and one light chain derived from the first antibody or the first antigen-binding region, and one heavy chain and one light chain derived from the second antibody or the second antigen-binding region. When both a monospecific antibody or its antigen-binding region recognize different epitopes on different antigens, the resulting heterodimer is a bispecific antibody.

[0219] In DuoBody technology, each monospecific antibody or antigen-binding region must contain a heavy chain constant region with a single point mutation in the CH3 domain. This point mutation enables stronger interactions between the CH3 domains of the resulting bispecific antibody than between any single CH3 domain of the monospecific antibody or antigen-binding region. The single point mutation in each monospecific antibody or antigen-binding region is located at residues 366, 368, 370, 399, 405, 407, or 409 of the heavy chain constant region's CH3 domain, numbered according to the EU numbering system, as described, for example, in International Publication No. 2011 / 131746. Furthermore, the single point mutation is located at a different residue in one monospecific antibody or antigen-binding region compared to the other monospecific antibody or antigen-binding region. For example, one monospecific antibody or antigen-binding region may contain mutation F405L (i.e., a mutation from phenylalanine to leucine at residue 405), while another monospecific antibody or antigen-binding region may contain mutation K409R (i.e., a mutation from lysine to arginine at residue 409), numbered according to the EU numbering system. The heavy chain constant region or antigen-binding region of a monospecific antibody may be an IgG1, IgG2, IgG3, or IgG4 isotype (e.g., human IgG1 isotype), and the bispecific antibody produced by DuoBody technology may retain Fc-mediated effector function.

[0220] Another method for generating bispecific antibodies is called the “knob-into-hole” strategy (see, for example, International Publication 2006 / 028936). This technique reduces mispairing of Ig heavy chains by mutating selected amino acids that form the interface of the CH3 domain of IgG. At a position within the CH3 domain where the two heavy chains directly interact, an amino acid with a small side chain (hole) is introduced into the sequence of one heavy chain, and an amino acid with a large side chain (knob) is introduced into the position of the corresponding interacting residue on the other heavy chain. In some embodiments, the compositions of this disclosure have immunoglobulin chains in which the CH3 domain is modified by mutating selected amino acids that interact at the interface between two polypeptides in order to preferentially form bispecific antibodies. Bispecific antibodies may consist of immunoglobulin chains of the same subclass (e.g., IgG1 or IgG3) or different subclasses (e.g., IgG1 and IgG3, or IgG3 and IgG4).

[0221] Bispecific antibodies may, in some cases, contain heterodimers of IgG4 and IgG1, IgG4 and IgG2, IgG4 and IgG3, or IgG1 and IgG3 chains. Such heterodimeric chain antibodies can be routinely manipulated to promote heterodimeric chain formation, for example, by modifying selected amino acids that form the interface between the CH3 domains of human IgG4 and IgG1 or IgG3.

[0222] In certain embodiments, an antibody described herein that binds to the same PC epitope as the anti-PC antibody described herein is a human antibody. In certain embodiments, an antibody described herein that competitively blocks any one of the antibodies described herein from binding to PC (e.g., in a dose-dependent manner) is a human antibody. Human antibodies can be produced using any method known in the art. For example, a transgenic mouse that cannot express functional endogenous immunoglobulins but can express human immunoglobulin genes can be used. In particular, the human heavy-chain and light-chain immunoglobulin gene complex can be introduced into mouse embryonic stem cells randomly or by homologous recombination. Alternatively, the human variable region, constant region, and diversity region can be introduced into mouse embryonic stem cells in addition to the human heavy-chain and light-chain genes. The mouse heavy-chain and light-chain immunoglobulin genes can be made non-functional separately or simultaneously with the introduction of human immunoglobulin loci by homologous recombination. In particular, homozygous deletion of the JH region prevents endogenous antibody production. Modified embryonic stem cells are proliferated and microinjected into blastocysts to generate chimeric mice. These chimeric mice are then bred to produce homozygous offspring expressing human antibodies. Transgenic mice are immunized in the usual manner with all or part of a selected antigen, e.g., an antigen (e.g., PC). Antigen-directed monoclonal antibodies can be obtained from transgenic mice immunized using conventional hybridoma techniques. The human immunoglobulin transgenes present in transgenic mice are rearranged during B-cell differentiation, followed by class switching and somatic mutations. Therefore, it is possible to produce therapeutically useful IgG, IgA, IgM, and IgE antibodies using such techniques. For an overview of this technique for producing human antibodies, see Lonberg N & Huszar D (1995) Int Rev Immunol 13:65-93, which is incorporated herein by reference in its entirety.For a detailed description of this technique for generating human antibodies and human monoclonal antibodies, and of protocols for producing such antibodies, see, for example, International Publication Nos. 98 / 24893, 96 / 34096, and 96 / 33735, and U.S. Patents Nos. 5,413,923, 5,625,126, 5,633,425, 5,569,825, 5,661,016, 5,545,806, 5,814,318, and 5,939,598, all of which are incorporated herein by reference in their entirety. Examples of mice capable of producing human antibodies include XenoMouse® (Abgenix, Inc.; U.S. Patent Nos. 6,075,181 and 6,150,184), HuAb-Mouse® (Medarex, Inc. / Gen Pharm; U.S. Patent Nos. 5,545,806 and 5,569,825), TransChromo Mouse® (Kirin), and KM Mouse® (Medarex / Kirin), all of which are incorporated herein by reference in their entirety.

[0223] Human antibodies that specifically bind to PC can be prepared using antibody libraries derived from human immunoglobulin sequences by various methods known in the art, including the phage display method described above. See also U.S. Patents 4,444,887, 4,716,111, and 5,885,793, as well as International Publications 98 / 46645, 98 / 50433, 98 / 24893, 98 / 16654, 96 / 34096, 96 / 33735, and 91 / 10741, all of which are incorporated herein by reference in their entirety.

[0224] In certain embodiments, human antibodies can be produced using mouse-human hybridomas. For example, human peripheral blood lymphocytes transformed with Epstein-Barr virus (EBV) can be fused with mouse myeloma cells to produce mouse-human hybridomas that secrete human monoclonal antibodies, and these mouse-human hybridomas can be screened to determine which secrete human monoclonal antibodies that specifically bind to a target antigen (e.g., PC). Such methods are known and described in the art; see, for example, Shinmoto H et al., (2004) Cytotechnology 46:19-23 and Naganawa Y et al., (2005) Human Antibodies 14:27-31, each of which is incorporated herein by reference in whole.

[0225] kit Furthermore, kits comprising one or more antibodies described herein, or their pharmaceutical compositions or conjugates, are also provided. In specific embodiments, pharmaceutical packs or kits are provided comprising one or more containers filled with one or more components of the pharmaceutical compositions described herein, for example, one or more antibodies described herein. In certain embodiments, the kit comprises the pharmaceutical compositions described herein and any prophylactic or therapeutic agent as described herein. Optionally, accompanying such container(s) may be a notice in the form prescribed by a government agency regulating the manufacture, use, or sale of a pharmaceutical or biological product, the notice reflecting approval by the agency for manufacture, use, or sale for human administration.

[0226] Kits that can be used in the manner described above are also provided. In certain embodiments, the kit comprises one or more containers containing the antibodies described herein, preferably purified antibodies. In specific embodiments, the kit described herein comprises substantially isolated PC antigen as a control. In another specific embodiment, the kit described herein further comprises a control antibody that does not react with the PC antigen. In yet another specific embodiment, the kit described herein comprises one or more elements for detecting the binding of an antibody to the PC antigen (for example, the antibody can be conjugated to a detectable substrate such as a fluorescent compound, an enzyme substrate, a radioactive compound, or a luminescent compound, or a second antibody that recognizes a first antibody can be conjugated to a detectable substrate). In specific embodiments, the kit provided herein may include recombinantly produced or chemically synthesized PC antigens. The PC antigen included in the kit can also be conjugated to a solid support. In a more specific embodiment, the detection means of the kit comprises a solid support to which the PC antigen is bound. Such a kit may also include an unbound reporter-labeled anti-human antibody or an anti-mouse / rat antibody. In this embodiment, the binding of an antibody to the PC antigen can be detected by the binding of the reporter-labeled antibody. In certain embodiments, the Disclosure relates to the use of the Kits of the Disclosure for in vitro assays and / or detection of PC antigens in biological samples. [Examples]

[0227] The following examples are provided for illustrative purposes only and are not limiting.

[0228] Example 1: Screening Overview A multifactor optimization platform was used for antibody optimization. This platform was designed to optimize affinity, burden removal, thermal stability, interspecies cross-reactivity, and selective activation.

[0229] E06 variable weight (VH) and variable kappa (VK) CDR sequences were grafted into four fully human VH germlines (VH1-46, VH3-15, VH3-23, VH5-51) and four fully human VK germlines (VK1-39, VK2-28, VK3-15, VK4-1), respectively. Of the 16 combinations created, two were selected for platform library construction: E06_VK1-39 / VH3-23 and E06_VK4-1 / VH3-23. The individual VH and VK sequences are shown in Table 2. [Table 2]

[0230] Phage display libraries of reformatted scFv framework grafts were designed based on structural and sequence analysis to retain CDR residues identified to exhibit critical target-binding interactions. The remaining CDR residues were used in an induced three-directional sequence space search to maximize the tolerance of human residues while limiting potential impacts on target binding. Firstly, CDRH1, CDRH2, CDRL1, CDRL2, and CDRL3 were modified from parental CDRs to human germline CDRs, with up to four changes per CDR. Secondly, CDRH1, CDRH2, CDRL1, CDRL2, and CDRL3 were modified from germline to parental CDR residues, with up to four changes. Thirdly, all CDRs were subjected to a single-amino acid NNK walk, omitting residue changes that would result in biochemical burden. Combining the E06 VK1-39 / VH3-23 and VK4-1 / VH3-23 scFv libraries resulted in 4 × 10⁶ 9 This represents the total size of the individual transformers (referred to as "platform libraries").

[0231] Phosphocholine (PC) conjugate peptides were used for ELISA-based phage panning and primary scFv screening. A summary of the PC conjugate peptides used is shown in Table 3. [Table 3]

[0232] Phage-peptide complexes were captured on peptide biotin tags with streptavidin-coated beads. After washing, the bound phages were eluted with triethylamine and titrated to determine concentration. After panning in each round, the eluted phages were used to infect with E. coli ER2738 for the proliferation of phage particles used in subsequent round selections.

[0233] Single colonies (380 colonies from each, totaling 1140 colonies) were cultured from the panning arms of Round 4, and scFv was Sanger sequenced. 736 unique sequences were rearranged on 96-well plates and proceeded to primary screening by scFv ELISA with peptides 1, 4, 5, and 8.

[0234] A total of 368 unique conjugates for peptide 5 and / or peptide 1 from the primary screening were rearranged and proceeded to secondary screening by ELISA (n=2) on peptides 1, 4, 5, and 8, as well as BSA. A total of 257 unique scFvs were identified that showed good reproducibility between the primary and secondary screening data, and between replicates. These 257 sequences specifically bound to at least one of peptides 1 and 5, but not to peptide 4, peptide 8, or BSA (the positive OD450 threshold was determined using the mean OD450 + 3 × SD of the negative control (PD-1 specific scFv): see Example 2 for the method).

[0235] Example 2: Overview of scFv ELISA Periplasm extract (PPE) containing scFv was generated by osmotic shock. 5-10 μl of glycerol stock was inoculated into starter cultures (2×YT containing 2% glucose, tetracycline (15 μg / mL), and carbenicillin (100 μg / mL), 700 μl / well in deep-well plates) and grown overnight at 30°C and 700 rpm. The following day, 30 μl of starter culture was inoculated into expression cultures (2×YT containing 0.1% glucose and carbenicillin (100 μg / mL), 1 ml / well in deep-well plates) and grown to OD600 0.5 at 37°C and 700 rpm. The cultures were induced with 1 mM IPTG and grown for 6 hours at 25°C and 700 rpm.

[0236] After 6 hours, the plate was centrifuged to pelletize the bacteria and stored overnight at -20°C. The following day, the pellet was thawed and resuspended in 80 μl of periplasm extraction buffer containing a protease inhibitor, and placed on a shaker at 4°C and 500 rpm for 10 minutes. After 10 minutes, 240 μl of water and the protease inhibitor were added to each well and incubated at 4°C and 500 rpm with shaking for 1 hour. The plate was then centrifuged at 4300 × g at 4°C for 10 minutes. The supernatant was passed through a filter plate and stored at -80°C.

[0237] For scFv-binding ELISA analysis, 96-well polystyrene plates were coated overnight at 4°C with peptides 1, 4, 5, 8, or 100 μL of BSA (10 μg / mL) in PBS. The plates were washed three times with PBS-Tween and blocked at room temperature for 1 hour with 100 μL of 1% BSA-TBS. 96-well streptavidin-coated plates were coated at room temperature for 1 hour with 100 μL of biotinylated PC-peptide (1 μg / mL) in PBS.

[0238] After washing all plates three times with PBS-Tween, 40 μL of aliquot PPE was added to the assay plate along with 50 μL of 10% BSA, and the plate was incubated at room temperature for 1 hour. After washing, 100 μL of HRP conjugate anti-V5 in 5% BSA was added, and the plate was incubated at room temperature for 30 minutes. The plate was washed, and 100 μL of TMB substrate was added. After color development at room temperature, the reaction was stopped with 100 μL of 1N HCl. The amount of scFv bound to each well was quantified by absorbance measurement at 450 nm (OD450). The binding data for 257 unique scFv identified in Example 1 are summarized in Table 4. [Table 4] TIFF2026509512000010.tif253170TIFF2026509512000011.tif248170TIFF20265095120 00012.tif233170TIFF2026509512000013.tif248170TIFF2026509512000014.tif159170

[0239] Example 3: Overview of IgG ELISA Forty-eight scFv cells were selected and reformatted to human IgG1 with parental E06 antibody using standard techniques. For antibody-binding ELISA analysis, a 384-well polystyrene microtiter plate was coated with 20 μL of antigen (5 μg / mL PC-BSA) in PBS at room temperature for 2 hours. The plate was washed three times with DELFIA® washing buffer and then blocked with 20 μL / well of 1% BSA-TBS at room temperature for 1 hour. After washing, 15 μL of titrated antibody in 1% BSA-TBS was added to the wells and incubated at room temperature for 1 hour. After further washing, 15 μL / well of DELFIA® Eu-N1 anti-human IgG secondary antibody in TBS was added to the wells and incubated at room temperature for 1 hour. The plate was washed again and incubated with DELFIA® enhancement buffer at room temperature for 5 minutes, after which the amount of antibody bound to each well was quantified by TRF measurement. Table 5 summarizes the binding data for the parent E06 IgG1 antibody and various IgG1 conjugates. No binding to BSA was detected for any of the antibodies shown in Table 5.

[0240] The physical properties of the antibodies (melting temperature, aggregation temperature, and polydispersity index) were determined using Uncle instruments (Unchained Labs). The physical properties of the parent E06 IgG1 antibody and various IgG1 conjugates are summarized in Table 5. [Table 5] TIFF2026509512000016.tif57170

[0241] Example 4: Further characterization of IgG The selected human IgG1 antibodies (see Example 3) were further analyzed for antibody yield, stress testing, and immunogenicity. Yield was determined by transfection of HEK293 cells and subsequent expression in 25 mL cultures. For some antibodies, yield was determined in scaled-up 40 L transient transfection cultures. Antibody products were purified using protein A, and the amount of antibody obtained was measured by absorbance at 280 nm (OD280). The yield values ​​for the parent E06 IgG1 antibody and selected IgG1 conjugates are shown in Table 6. [Table 6]

[0242] Parental E06 IgG1 antibody and human IgG1 PcOxP_DB01_H08 antibody were analyzed in forced degradation studies to compare the relative stability and stress resistance between the antibodies. Samples of each antibody were subjected to temperature stress, oxidative stress, and low and high pH stress, and the samples were analyzed and compared with unstressed samples. Samples were analyzed using various analytical techniques. In addition to concentration measurements and visual inspection, chemical stability was assessed by protein-level RPC-MS at intact and reduced antibody levels, peptide mapping analysis under reducing conditions, iCIEF analysis, and CGE-SDS under non-reducing and reducing conditions, while physical stability was assessed using SE-HPLC.

[0243] For temperature stress, samples of each antibody were stored at 40°C for 2, 3, or 4 weeks. For low pH stress, the pH of each antibody sample was adjusted to 3.2 using 0.12 M HCl, the sample was incubated at 25°C for 2 hours, and then the buffer was returned to the formulation buffer. For high pH stress (forced deamide), the pH of each antibody sample was adjusted to 9.0 using 0.5 M Tris pH11, the sample was incubated at 37°C for 3 days, and then the buffer was returned to the formulation buffer. For oxidative stress, a 3% H2O2 solution was added to each antibody sample to reach a final concentration of 0.03%, the sample was incubated at 25°C for 1 day, and then the buffer was returned to the formulation buffer. The results of the stress tests and stability assays are qualitatively summarized in Table 7 ("+": moderately stable; "++": very stable). [Table 7]

[0244] In vivo immunogenicity for parental E06 IgG1 antibody and human IgG1 PcOxP_DB01_H08 and PcOxP_DB03_C03 antibodies was predicted using the in vitro Epibase® immunogenicity assay (Lonza Biologics), the in silico Epibase® platform (Lonza Biologics), and the in silico ISPRI® platform (EpiVax, Inc.). The immunogenicity profiles are summarized in Table 8. Overall, the immunogenicity assays indicate that PcOxP_DB01_H08 and PcOxP_DB03_C03 have lower immunogenicity risk and favorable profiles compared to several established antibody drugs, respectively. [Table 8] [Table 9] TIFF2026509512000021.tif232170TIFF2026509512000022.tif251170TIFF2026509512000023.tif243170TIFF2026509512000024.tif242170TIFF2026509512000025.tif234170TIFF2026509512000026.tif233170TIFF2026509512000027.tif239170TIFF2026509512000028.tif237170TIFF2026509512000029.tif247170TIFF2026509512000030.tif243170TIFF2026509512000031.tif238170TIFF2026509512000032.tif228170TIFF2026509512000033.tif238170TIFF2026509512000034.tif236170TIFF2026509512000035.tif225170TIFF2026509512000036.tif239170TIFF2026509512000037.tif245170TIFF2026509512000038.tif242170TIFF2026509512000039.tif237170TIFF2026509512000040.tif233170TIFF2026509512000041.tif247170TIFF2026509512000042.tif237170TIFF2026509512000043.tif240170TIFF2026509512000044.tif230170TIFF2026509512000045.tif236170TIFF2026509512000046.tif231170TIFF2026509512000047.tif234170TIFF2026509512000048.tif235170TIFF2026509512000049.tif223170TIFF2026509512000050.tif214170

[0245] The present invention is not limited in scope by the specific embodiments described herein. In fact, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

[0246] All references cited herein (e.g., publications, patents, or patent applications) are incorporated herein by reference in their entirety to the same extent that each individual reference (e.g., publications, patents, or patent applications) is specifically and individually indicated to be incorporated in its entirety by reference for any purpose.

[0247] Other embodiments are within the scope of the following claims.

Claims

1. An antibody that specifically binds to phosphocholine (PC), wherein the antibody comprises VH, which includes one CDRH1, CDRH2, and CDRH3 amino acid sequence from any one of the VH amino acid sequences described in SEQ ID NOs: 1 to 180, and VL, which includes one CDRL1, CDRL2, and CDRL3 amino acid sequence from any one of the VL amino acid sequences described in SEQ ID NOs: 181 to 368.

2. The antibodies are, respectively, sequence numbers 1 and 181; 2 and 182; 3 and 183; 4 and 184; 5 and 185; 1 and 184; 6 and 184; 1 and 186; 7 and 187; 8 and 188; 9 and 189; 10 and 190; 4 and 191; 11 and 192; 12 and 193; 13 and 194; 14 and 195; 15 and 196; 16 and 184; 17 and 197; 18 and 198; 1 and 199; 19 and 200; 20 and 201; 21 and 202; 22 and 203; 23 and 204; 24 and 205;25 and 184;4 and 206;26 and 207;27 and 208;28 and 209;9 and 210;29 and 211;30 and 212;31 and 213;32 and 214;33 and 215;34 and 216;17 and 217;1 and 218;35 and 184;36 and 184;35 and 219;35 and 220;37 and 184;38 and 221;39 and 222;9 and 223;9 and 224;40 and 184;41 and 225;8 and 226;42 and 227;35 and 228; 43 and 229; 4 and 230; 9 and 231; 8 and 232; 1 and 233; 8 and 234; 44 and 184; 1 and 235; 45 and 236; 30 and 237; 46 and 184; 47 and 238; 48 and 239; 1 and 230; 49 and 190; 44 and 240; 50 and 184; 51 and 241; 52 and 242; 53 and 243; 54 and 184; 55 and 244; 56 and 245; 57 and 246; 58 and 247; 59 and 248; 60 and 249; 8 and 250; 61 and 184; 35 and 251; 62 and 252; 1 and 253; 63 and 254; 64 and 181; 65 and 255; 66 and 256; 67 and 257; 68 and 198; 35 and 258; 69 and 259; 70 and 260; 71 and 261; 72 and 262; 73 and 263; 74 and 264; 75 and 265; 76 and 266; 77 and 267; 4 and 205; 78 and 268; 1 and 269; 27 and 269; 79 and 269; 80 and 270; 81 and 269; 22 and 269;82 and 269; 83 and 271; 84 and 272; 1 and 273; 8 and 274; 8 and 275; 22 and 276; 8 and 277; 85 and 278; 86 and 279; 87 and 269; 88 and 280; 89 and 269; 60 and 281; 90 and 269; 91 and 282; 22 and 283; 92 and 269; 93 and 284; 94 and 285; 95 and 286; 96 and 269; 73 and 287; 97 and 288; 68 and 289; 98 and 269; 99 and 290; 100 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304; 72 and 305; 114 and 306; 115 and 307; 116 and 269; 117 and 308; 118 and 269; 110 and 309; 119 and 310; 120 and 311; 121 and 269; 122 and 312; 123 and 289; 9 and 313; 124 and 314; 120 and 289; 125 and 315; 110 and 316; 126 and 317; 127 and 318; 49 and 319; 128 and 320; 9 and 321; 22 and 322; 129 and 323; 130 and 269; 131 and 324; 132 and 325; 133 and 326; 134 and 327; 53 and 328 ;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340;149 and 341;150 and 342;151 and 343;152 and 344;94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 356; 166 and 357; 167 and 358; 110 and 334; 87 and 359; 168 and 36 The antibody according to claim 1, comprising the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences of the VH and VL amino acid sequences described in 0;169 and 269;170 and 269;171 and 269;172 and 361;8 and 362;173 and 304;174 and 269;1 and 363;175 and 364;176 and 365;177 and 269;178 and 366;179 and 269;180 and 367; or 89 and 368.

3. The antibodies are, respectively, sequence numbers 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381, 428 , and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 484; 385, 428, and 484; 372, 438, and 484; 388, 428, and 494; 369, 439, and 484; 389, 428, and 484; 381, 428, and 484; 376, 440, and 495; 390, 441, and 484; 370, 442, and 484; 391, 428, and 484; 392, 432, and 484; 377, 443, and 484; 393, 428, and 484; 383, 444, and 489; 394, 428, and 484; 383, 444, and 496; 376, 434, and 484; 395, 445, and 497; 396, 446, and 487; 392, 428 , and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484; 402, 436, and 484; 381, 451, and 484; 403, 428, and 484;369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428, and 497; 403, 453, and 484; 402, 454, and 484; 383, 434 , and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428, and 484; 415, 428, and 489; 369, 428, and 491; 403, 458 , and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512; 381, 428, and 493; 383, 470, and 501; 404, 471, and 484;418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515; 376, 428, and 504; 422, 469, and 484; 3 69, 428, and 516; 405, 460, and 484; 413, 431, and 484; 406, 429, and 484; 383, 428, and 517; 377, 437, and 518; 369, 477, and 487; 403, 464, and 484; 369, 454, and 484; 372, 478, and 519; 423, 429, and 484; 382, ​​434, and 484; 424, 479, and 484; 376 , 429, and 484; 425, 428, and 484; 403, 428, and 520; 417, 460, and 484; 409, 460, and 484; 404, 452, and 484; 383, 428, and 521; 372, 480, and 493; 369, 477, and 484; 369, 481, and 484; 384, 452, and 484; 377, 482, and 484; 410, 483, and 484; 384, The antibody according to claim 1 or 2, comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 428, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or 400, 452, and 484.

4. The antibodies are, respectively, sequence numbers 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523, 601 , and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 661; 523, 610, and 659; 523, 611, and 668; 523, 611, and 658; 523, 596, and 662; 523, 612, and 667; 523, 613, and 668; 523, 591, and 669; 523, 591, and 663; 523, 614, and 659; 532, 615, and 662; 523, 616, and 670; 523, 591, and 668; 523, 617, and 671; 533, 595, and 659; 523, 618, and 659; 534, 619, and 662; 535, 620, and 659; 536, 621, and 659; 537, 603, and 671; 523, 622 , and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671; 539, 591, and 662; 523, 591, and 670; 523, 611, and 659;537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610, and 674; 523, 634, and 671; 545, 591, and 661; 523, 635 , and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616, and 660; 523, 602, and 677; 523, 616, and 659; 523, 630 , and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678; 571, 630, and 663; 572, 591, and 664; 573, 591, and 670;523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 523, 595, and 659; 523, 600, and 659; 523, 649, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 591, and 659; 584, 591, and 671; 523, 5 91, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; 560, 591, and 659; The antibody according to any one of the prior claims, comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or 523, 606, and 680.

5. The antibodies are sequence numbers 369, 428, 484, 523, 591, and 656; 370, 429, 485, 523, 592, and 657; 371, 428, 486, 523, 593, and 658; 372, 428, 484, 523, 591, and 659; 373, 429, 484, 523, 594, and 659; 369, 428, 484, 523, 591, and 659; 374, 430, 484, 523, 591, and 659; 369, 428, 484, 523, 595, and 660; 375, 428, 487, 523, 596 , and 659; 376, 428, 484, 524, 591, and 659; 377, 428, 484, 523, 591, and 661; 378, 428, 484, 523, 591, and 662; 372, 428, 484, 523, 597, and 658; 379, 428, 484, 523, 598, and 662; 372, 428, 488, 525, 599, and 659; 376, 431, 484, 523, 600, and 663; 380, 432, 484, 523, 591, and 664; 372, 428, 489, 523, 601, and 659; 381, 428, 490, 523, 591, and 659; 382, ​​428, 484, 526, 597, and 665; 381, 428, 491, 523, 602, and 659; 369, 428, 484, 527, 591, and 661; 377, 428, 492, 528, 591, and 662; 376, 433, 484, 529, 603, and 662; 383, 434, 484, 523, 604, and 666; 383, 428, 484, 523, 605, and 667; 384, 435, 484, 530, 606, and 6 59; 369, 436, 484, 523, 607, and 659; 372, 437, 487, 523, 591, and 659; 372, 428, 484, 523, 608, and 659; 385, 428, 493, 523, 609, and 662; 386, 428, 484, 531, 591, and 663; 372, 437, 484, 523, 608, and 661; 377, 428, 484, 523, 610, and 659; 387, 428, 484, 523, 611, and 668; 385, 428, 484, 523, 611, and 658;372, 438, 484, 523, 596, and 662; 388, 428, 494, 523, 612, and 667; 369, 439, 484, 523, 613, and 668; 389, 428, 484, 523, 591, and 669; 382, ​​428, 484, 523, 591, and 663; 369, 428, 484, 523, 614, and 659; 381, 428, 484, 523, 591, and 659; 376, 440, 495, 523, 591, and 659; 381, 428, 484, 532, 615, and 662; 381, 42 8, 484, 523, 616, and 670; 390, 441, 484, 523, 591, and 659; 370, 442, 484, 523, 591, and 668; 391, 428, 484, 523, 617, and 671; 377, 428, 484, 533, 595, and 659; 377, 428, 484, 523, 618, and 659; 392, 432, 484, 523, 591, and 659; 377, 443, 484, 534, 619, and 662; 376, 428, 484, 535, 620, and 659; 393, 428, 484, 5 36, 621, and 659; 381, 428, 484, 537, 603, and 671; 383, 444, 489, 523, 622, and 59; 372, 428, 484, 523, 617, and 659; 377, 428, 484, 523, 623, and 659; 376, 428, 484, 538, 607, and 672; 369, 428, 484, 523, 616, and 671; 376, 428, 484, 523, 624, and 659; 394, 428, 484, 523, 591, and 659; 369, 428, 484, 523, 620, and 659; 383, 444, 496, 523, 625, and 660; 385, 428, 484, 523, 602, and 671; 376, 434, 484, 523, 591, and 659; 395, 445, 497, 523, 611, and 663; 396, 446, 487, 523, 626, and 670; 369, 428, 484, 523, 617, and 659; 392, 428, 484, 523, 591, and 662; 394, 428, 484, 523, 606, and 673; 393, 428, 498, 523, 591, and 659;376, 428, 495, 523, 627, and 671; 369, 440, 484, 523, 628, and 671; 397, 428, 484, 539, 591, and 662; 389, 447, 484, 523, 591, and 659; 398, 448, 492, 523, 591, and 670; 373, 449, 490, 523, 611, and 659; 399, 428, 484, 537, 613, and 671; 400, 450, 484, 540, 598, and 659; 401, 428, 484, 541, 591, and 659; 390, 42 8, 484, 542, 629, and 659; 376, 428, 484, 523, 591, and 671; 400, 428, 499, 523, 591, and 659; 381, 428, 484, 523, 630, and 667; 384, 429, 484, 543, 631, and 659; 369, 428, 484, 523, 632, and 664; 402, 436, 484, 523, 597, and 661; 381, 451, 484, 523, 591, and 656; 403, 428, 484, 523, 633, and 659; 369, 429, 484, 5 44, 607, and 659; 404, 429, 484, 523, 630, and 659; 372, 429, 484, 523, 602, and 659; 381, 428, 484, 523, 591, and 665; 388, 428, 484, 523, 610, and 674; 405, 451, 484, 523, 634, and 671; 406, 428, 484, 545, 591, and 661; 400, 428, 484, 523, 635, and 667; 404, 428, 484, 546, 591, and 659; 384, 432, 500, 523, 636, and 664; 376, 452, 501, 547, 591, and 663; 407, 428, 484, 548, 619, and 659; 375, 428, 484, 523, 637, and 659; 372, 428, 484, 523, 607, and 659; 408, 428, 484, 523, 638, and 659; 386, 428, 484, 523, 591, and 659; 369, 428, 497, 523, 591, and 659; 403, 453, 484, 549, 591, and 659; 402, 454, 484, 523, 591, and 659;383, 428, 484, 523, 591, and 659; 383, 434, 500, 523, 591, and 659; 376, 455, 502, 550, 591, and 675; 383, 435, 484, 523, 591, and 664; 369, 428, 484, 523, 602, and 659; 376, 428, 484, 551, 591, and 659; 376, 428, 484, 552, 591, and 659; 383, 428, 484, 553, 591, and 676; 376, 428, 484, 554, 591, and 659; 376, 42 8, 490, 523, 591, and 676; 369, 428, 503, 555, 606, and 659; 409, 428, 484, 523, 591, and 659; 383, 428, 504, 556, 591, and 659; 369, 434, 484, 523, 591, and 659; 390, 428, 484, 523, 637, and 677; 369, 456, 484, 523, 591, and 659; 370, 457, 484, 557, 591, and 659; 383, 428, 484, 558, 591, and 659; 404, 458, 499, 5 23, 591, and 659; 410, 428, 484, 523, 591, and 677; 383, 459, 484, 553, 639, and 659; 407, 428, 489, 549, 591, and 667; 374, 434, 484, 523, 591, and 659; 404, 428, 484, 559, 617, and 666; 411, 460, 505, 523, 591, and 672; 372, 429, 484, 523, 606, and 659; 383, 428, 485, 523, 591, and 659; 386, 460, 490, 523, 601, and 659; 412, 455, 493, 523, 592, and 670; 372, 428, 484, 560, 591, and 667; 372, 428, 484, 523, 606, and 659; 383, 461, 484, 523, 591, and 677; 369, 429, 484, 523, 606, and 659; 369, 434, 484, 530, 591, and 659; 376, 437, 484, 561, 603, and 666; 383, 462, 500, 523, 591, and 659; 383, 428, 484, 562, 591, and 659;400, 463, 484, 523, 616, and 660; 376, 464, 484, 523, 602, and 677; 400, 434, 506, 549, 591, and 659; 369, 436, 507, 523, 591, and 659; 404, 429, 484, 523, 591, and 659; 384, 429, 484, 523, 616, and 659; 369, 429, 484, 523, 591, and 659; 377, 465, 484, 523, 630, and 660; 413, 459, 484, 563, 591, and 659; 369, 45 2, 484, 523, 591, and 663; 414, 466, 484, 523, 640, and 659; 373, 428, 484, 523, 591, and 659; 415, 428, 489, 564, 591, and 659; 369, 428, 484, 523, 591, and 671; 400, 428, 484, 523, 641, and 677; 369, 428, 491, 565, 591, and 662; 403, 458, 508, 523, 642, and 664; 392, 467, 484, 523, 591, and 659; 369, 452, 509, 5 66, 643, and 659; 388, 468, 490, 523, 591, and 659; 369, 452, 484, 523, 644, and 667; 377, 440, 484, 567, 606, and 659; 370, 428, 484, 523, 591, and 656; 384, 467, 487, 523, 591, and 659; 392, 469, 510, 568, 591, and 659; 378, 464, 484, 523, 606, and 659; 377, 428, 484, 569, 645, and 659; 416, 434, 484, 523, 603, and 659; 370, 428, 484, 523, 606, and 659; 370, 428, 511, 570, 628, and 678; 369, 452, 484, 571, 630, and 663; 376, 428, 502, 572, 591, and 664; 417, 428, 484, 523, 591, and 665; 392, 428, 484, 573, 591, and 670; 376, 428, 512, 523, 646, and 659; 377, 428, 484, 574, 591, and 670; 383, 428, 484, 523, 647, and 667;381, 428, 493, 575, 591, and 659; 383, 470, 501, 523, 591, and 659; 404, 471, 484, 537, 591, and 659; 418, 428, 484, 523, 635, and 670; 383, 472, 513, 576, 591, and 659; 419, 428,; 484, 577, 606, and 659; 397, 428, 484, 523, 648, and 679; 372, 473, 484, 523, 591, and 664; 383, 474, 484, 523, 592, and 659; 372, 428, 484, 523, 591, and 663; 420, 434, 484, 523, 595, and 659; 370, 428, 514, 523, 591, and 659; 369, 428, 484, 523, 591, and 677; 383, 428, 484, 523, 600, and 659; 369, 475, 484, 523, 591, and 659; 421, 436, 484, 523, 649, and 664; 378, 452, 484, 523, 591, and 659; 376, 476, 515, 578, 641, and 659; 376, 428, 504, 523, 591, and 667; 422, 469, 484, 523, 606, and 660; 383, 461, 484, 523, 616, and 659; 369, 428, 484, 523, 650, and 656; 369, 428, 516, 523, 591, and 659; 405, 460, 484, 523, 6 18, and 659; 413, 431, 484, 523, 591, and 659; 382, ​​428, 484, 579, 621, and 671; 383, 428, 484, 523, 646, and 659; 372, 428, 484, 580, 591, and 660; 406, 429, 484, 527, 591, and 659; 383, 428, 517, 581, 595, and 659; 377, 437, 518, 582, 640, and 664; 369, 477, 487, 583, 651, and 659; 403, 464, 484, 534, 591, and 659; 383, 459, 484, 523, 606, and 659; 369, 454, 484, 584, 591, and 671; 372, 478, 519, 523, 591, and 680; 423, 429, 484, 523, 591, and 681; 383, 428, 484, 585, 591, and 659; 369, 434, 484, 557, 591, and 659; 382, ​​434, 484, 523, 649, and 659; 424, 479, 484, 523, 616, and 660; 376, 429, 484, 586, 591, and 666;425, 428, 484, 523, 592, and 659; 369, 436, 484, 587, 652, and 659; 403, 428, 520, 588, 638, and 659; 417, 460, 484, 523, 591, and 659; 409, 460, 484, 523, 623, and 664; 404, 452, 484, 523, 591, and 659; 369, 428, 484, 523, 591, and 682; 383, 428, 521, 553, 653, and 659; 372, 4 80, 493, 523, 591, and 659; 369, 428, 484, 523, 654, and 659; 369, 477, 484, 523, 648, and 659; 369, 481, 484, 560, 591, and 659; 369, 452, 484, 523, 591, and 667; 409, 428, 484, 580, 606, and 659; 384, 452, 484, 589, 593, and 661; 377, 482, 484, 523, 591, and 659; 410, 483, 48 4, 523, 591, and 659; 384, 428, 489, 523, 591, and 659; 426, 429, 484, 523, 622, and 659; 376, 428, 484, 523, 637, and 676; 427, 428, 484, 523, 591, and 671; 373, 452, 506, 523, 591, and 659; 369, 428, 484, 590, 622, and 659; 369, 429, 521, 555, 591, and 661; 405, 428, 522, 523 The antibody according to any one of the prior claims, comprising the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences as described in 615, and 672; 376, 428, 510, 523, 591, and 659; 384, 428, 484, 523, 626, and 668; 410, 429, 484, 523, 591, and 659; 400, 452, 484, 523, 655, and 659; or 369, 434, 484, 523, 606, and 680.

6. The antibody according to any one of the prior claims, wherein the antibody comprises one of the VH amino acid sequences from SEQ ID NOs: 1 to 180.

7. The antibody according to any one of the prior claims, wherein the antibody comprises one of the VL amino acid sequences from SEQ ID NOs: 181 to 368.

8. The aforementioned VH and VL are, respectively, sequence numbers 1 and 181; 2 and 182; 3 and 183; 4 and 184; 5 and 185; 1 and 184; 6 and 184; 1 and 186; 7 and 187; 8 and 188; 9 and 189; 10 and 190; 4 and 191; 11 and 192; 12 and 193; 13 and 194; 14 and 195; 15 and 196; 16 and 184; 17 and 197; 18 and 198; 1 and 199; 19 and 200; 20 and 201; 21 and 202; 22 and 203; 23 and 204 ;24 and 205;25 and 184;4 and 206;26 and 207;27 and 208;28 and 209;9 and 210;29 and 211;30 and 212;31 and 213;32 and 214;33 and 215;34 and 216;17 and 217;1 and 218;35 and 184;36 and 184;35 and 219;35 and 220;37 and 184;38 and 221;39 and 222;9 and 223;9 and 224;40 and 184;41 and 225;8 and 226;42 and 227;35 and 228; 43 and 229; 4 and 230; 9 and 231; 8 and 232; 1 and 233; 8 and 234; 44 and 184; 1 and 235; 45 and 236; 30 and 237; 46 and 184; 47 and 238; 48 and 239; 1 and 230; 49 and 190; 44 and 240; 50 and 184; 51 and 241; 52 and 242; 53 and 243; 54 and 184; 55 and 244; 56 and 245; 57 and 246; 58 and 247; 59 and 248; 60 and 249; 8 and 250 ;61 and 184;35 and 251;62 and 252;1 and 253;63 and 254;64 and 181;65 and 255;66 and 256;67 and 257;68 and 198;35 and 258;69 and 259;70 and 260;71 and 261;72 and 262;73 and 263;74 and 264;75 and 265;76 and 266;77 and 267;4 and 205;78 and 268;1 and 269;27 and 269;79 and 269;80 and 270;81 and 269;22 and 269;82 and 269; 83 and 271; 84 and 272; 1 and 273; 8 and 274; 8 and 275; 22 and 276; 8 and 277; 85 and 278; 86 and 279; 87 and 269; 88 and 280; 89 and 269; 60 and 281; 90 and 269; 91 and 282; 22 and 283; 92 and 269; 93 and 284; 94 and 285; 95 and 286; 96 and 269; 73 and 287; 97 and 288; 68 and 289; 98 and 269; 99 and 290; 100 and 291; 4 and 292; 4 and 289; 101 and 284; 66 and 289; 89 and 293; 102 and 294; 103 and 269; 22 and 295; 104 and 296; 105 and 297; 106 and 270; 107 and 269; 67 and 269; 62 and 298; 66 and 269; 108 and 299; 109 and 300; 110 and 301; 111 and 302; 112 and 269; 113 and 303; 1 and 304; 72 and 305; 114 and 306; 115 and 307; 116 and 269; 117 and 308; 118 and 269; 110 and 309; 119 and 310; 120 and 311; 121 and 269; 122 and 312; 123 and 289; 9 and 313; 124 and 314; 120 and 289; 125 and 315; 110 and 316; 126 and 317; 127 and 318; 49 and 319; 128 and 320; 9 and 321; 22 and 322; 129 and 323; 130 and 269; 131 and 324; 132 and 325; 133 and 326; 134 and 327; 53 and 328 ;135 and 272;136 and 329;4 and 301;137 and 330;138 and 269;1 and 284;22 and 331;139 and 269;140 and 332;141 and 269;142 and 333;143 and 334;144 and 335;101 and 298;1 and 336;145 and 269;146 and 337;147 and 269;17 and 338;22 and 320;4 and 339;148 and 340;149 and 341;150 and 342;151 and 343;152 and 344;94 and 289; 153 and 345; 154 and 346; 155 and 347; 22 and 348; 89 and 282; 156 and 349; 157 and 296; 158 and 350; 159 and 329; 24 and 351; 160 and 352; 161 and 269; 162 and 353; 163 and 269; 1 and 354; 164 and 355; 165 and 269; 1 and 356; 166 and 357; 167 and 358; 110 An antibody according to any one of the prior claims, comprising the amino acid sequence described in 334; 87 and 359; 168 and 360; 169 and 269; 170 and 269; 171 and 269; 172 and 361; 8 and 362; 173 and 304; 174 and 269; 1 and 363; 175 and 364; 176 and 365; 177 and 269; 178 and 366; 179 and 269; 180 and 367; or 89 and 368.

9. The antibody according to any one of the prior claims, wherein the antibody is a single-stranded variable fragment (scFv).

10. The aforementioned antibody is optionally selected from human IgG １ , IgG ２ , IgG ３ , IgG ４ IgA １ , and IgA ２ An antibody according to any one of claims 1 to 8, comprising a heavy chain constant region or its Fc region, selected from the group consisting of the following.

11. The heavy chain constant region is human IgG １ The antibody according to claim 10.

12. The heavy chain constant region is human IgG ４ The antibody according to claim 10.

13. Human IgG ４ The antibody according to claim 12, wherein the amino acid sequence of the heavy chain constant region is numbered according to the EU numbering system and contains P at position 228.

14. The antibody according to any one of claims 10 to 13, wherein the heavy chain constant region is a variant of the wild-type heavy chain constant region, and the variant heavy chain constant region binds to the Fc gamma receptor (FcγR) with a lower affinity than the wild-type heavy chain constant region binds to the Fc gamma receptor (FcγR).

15. The antibody according to claim 14, wherein the amino acid sequence of the heavy chain constant region comprises, in each case, A at position 234, A at position 235, A, Q, or G at position 297, or A or G at position 329, as numbered according to the EU numbering system.

16. The antibody according to claim 14, wherein the amino acid sequence of the heavy chain constant region comprises, in each case, A at positions 234 and 235, A at positions 234, 235 and 329, or A at positions 234 and 235 and G at position 329, as numbered according to the EU numbering system.

17. The antibody according to any one of claims 10 to 16, wherein the heavy chain constant region is a variant of the wild-type heavy chain constant region, and the variant heavy chain constant region has an increased affinity for human neonatal Fc receptor (FcRn) at pH 6 compared to the affinity of the wild-type heavy chain constant region for human FcRn at pH 6.

18. The antibody according to claim 17, wherein the amino acid sequence of the heavy chain constant region comprises, in each case, L and S at positions 428 and 434, respectively, and K, F, and Y at positions 433, 434, and 436, respectively, or Y, T, and E at positions 252, 254, and 256, respectively, as numbered according to the EU numbering system.

19. The antibody according to any one of claims 1 to 18, comprising a light chain constant region and optionally a human kappa or lambda constant region.

20. A polypeptide comprising a VH having one of the CDRH1, CDRH2, and CDRH3 amino acid sequences from the VH amino acid sequences described in SEQ ID NOs: 1 to 180.

21. The polypeptides are, respectively, sequence numbers 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381, 428, and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 484; 385, 428, and 484; 372, 438, and 484; 388, 428, and 49 4;369, 439, and 484;389, 428, and 484;381, 428, and 484;376, 440, and 495;390, 441, and 484;370, 442, and 484;391, 428, and 484;392, 432, and 484;377, 443, and 484;393, 428, and 484;383, 444, and 489;394, 428, and 484;383, 444, and 496;376, 434, and 484;395, 445, and 497;396, 446, and 487;392, 42 8, and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484; 402, 436, and 484; 381, 451, and 484; 403, 428, and 484;369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428, and 497; 403, 453, and 484; 402, 454, and 484; 383, 434 , and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428, and 484; 415, 428, and 489; 369, 428, and 491; 403, 458 , and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512; 381, 428, and 493; 383, 470, and 501; 404, 471, and 484;418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515; 376, 428, and 504; 422, 469, and 484; 3 69, 428, and 516; 405, 460, and 484; 413, 431, and 484; 406, 429, and 484; 383, 428, and 517; 377, 437, and 518; 369, 477, and 487; 403, 464, and 484; 369, 454, and 484; 372, 478, and 519; 423, 429, and 484; 382, ​​434, and 484; 424, 479, and 484; 376 , 429, and 484; 425, 428, and 484; 403, 428, and 520; 417, 460, and 484; 409, 460, and 484; 404, 452, and 484; 383, 428, and 521; 372, 480, and 493; 369, 477, and 484; 369, 481, and 484; 384, 452, and 484; 377, 482, and 484; 410, 483, and 484; 384, 4 The polypeptide according to claim 20, comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 28, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or 400, 452, and 484.

22. The aforementioned VHs are sequence numbers 369, 428, and 484; 370, 429, and 485; 371, 428, and 486; 372, 428, and 484; 373, 429, and 484; 374, 430, and 484; 375, 428, and 487; 376, 428, and 484; 377, 428, and 484; 378, 428, and 484; 379, 428, and 484; 372, 428, and 488; 376, 431, and 484; 380, 432, and 484; 372, 428, and 489; 381, 428 , and 490; 382, ​​428, and 484; 381, 428, and 491; 377, 428, and 492; 376, 433, and 484; 383, 434, and 484; 383, 428, and 484; 384, 435, and 484; 369, 436, and 484; 372, 437, and 487; 385, 428, and 493; 386, 428, and 484; 372, 437, and 484; 387, 428, and 484; 385, 428, and 484; 372, 438, and 484; 388, 428, and 494; 369, 439, and 484; 389, 428, and 484; 381, 428, and 484; 376, 440, and 495; 390, 441, and 484; 370, 442, and 484; 391, 428, and 484; 392, 432, and 484; 377, 443, and 484; 393, 428, and 484; 383, 444, and 489; 394, 428, and 484; 383, 444, and 496; 376, 434, and 484; 395, 445, and 497; 396, 446, and 487; 392, 428 , and 484; 393, 428, and 498; 376, 428, and 495; 369, 440, and 484; 397, 428, and 484; 389, 447, and 484; 398, 448, and 492; 373, 449, and 490; 399, 428, and 484; 400, 450, and 484; 401, 428, and 484; 390, 428, and 484; 400, 428, and 499; 384, 429, and 484; 402, 436, and 484; 381, 451, and 484; 403, 428, and 484;369, 429, and 484; 404, 429, and 484; 372, 429, and 484; 388, 428, and 484; 405, 451, and 484; 406, 428, and 484; 400, 428, and 484; 404, 428, and 484; 384, 432, and 500; 376, 452, and 501; 407, 428, and 484; 375, 428, and 484; 408, 428, and 484; 369, 428, and 497; 403, 453, and 484; 402, 454, and 484; 383, 434 , and 500; 376, 455, and 502; 383, 435, and 484; 376, 428, and 490; 369, 428, and 503; 409, 428, and 484; 383, 428, and 504; 369, 434, and 484; 369, 456, and 484; 370, 457, and 484; 404, 458, and 499; 410, 428, and 484; 383, 459, and 484; 407, 428, and 489; 374, 434, and 484; 411, 460, and 505; 383, 428, and 485; 386, 460, and 490; 412, 455, and 493; 383, 461, and 484; 376, 437, and 484; 383, 462, and 500; 400, 463, and 484; 376, 464, and 484; 400, 434, and 506; 369, 436, and 507; 377, 465, and 484; 413, 459, and 484; 369, 452, and 484; 414, 466, and 484; 373, 428, and 484; 415, 428, and 489; 369, 428, and 491; 403, 458 , and 508; 392, 467, and 484; 369, 452, and 509; 388, 468, and 490; 377, 440, and 484; 370, 428, and 484; 384, 467, and 487; 392, 469, and 510; 378, 464, and 484; 416, 434, and 484; 370, 428, and 511; 376, 428, and 502; 417, 428, and 484; 376, 428, and 512; 381, 428, and 493; 383, 470, and 501; 404, 471, and 484;418, 428, and 484; 383, 472, and 513; 419, 428, and 484; 372, 473, and 484; 383, 474, and 484; 420, 434, and 484; 370, 428, and 514; 369, 475, and 484; 421, 436, and 484; 378, 452, and 484; 376, 476, and 515; 376, 428, and 504; 422, 469, and 484; 3 69, 428, and 516; 405, 460, and 484; 413, 431, and 484; 406, 429, and 484; 383, 428, and 517; 377, 437, and 518; 369, 477, and 487; 403, 464, and 484; 369, 454, and 484; 372, 478, and 519; 423, 429, and 484; 382, ​​434, and 484; 424, 479, and 484; 376 , 429, and 484; 425, 428, and 484; 403, 428, and 520; 417, 460, and 484; 409, 460, and 484; 404, 452, and 484; 383, 428, and 521; 372, 480, and 493; 369, 477, and 484; 369, 481, and 484; 384, 452, and 484; 377, 482, and 484; 410, 483, and 484; 384, 4 The polypeptide according to claim 20, comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences described in 28, and 489; 426, 429, and 484; 427, 428, and 484; 373, 452, and 506; 369, 429, and 521; 405, 428, and 522; 376, 428, and 510; 384, 428, and 484; 410, 429, and 484; or 400, 452, and 484.

23. The polypeptide according to claim 20, wherein the VH comprises any one of the amino acid sequences described in SEQ ID NOs: 1 to 180.

24. A polypeptide comprising a VL containing one of the CDRL1, CDRL2, and CDRL3 amino acid sequences from the VL amino acid sequences described in SEQ ID NOs. 181 to 368.

25. The polypeptides are, respectively, sequence numbers 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523, 601, and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 661; 523, 610, and 659; 523, 611, and 668; 523, 611, and 65 8; 523, 596, and 662; 523, 612, and 667; 523, 613, and 668; 523, 591, and 669; 523, 591, and 663; 523, 614, and 659; 532, 615, and 662; 523, 616, and 670; 523, 591, and 668; 523, 617, and 671; 533, 595, and 659; 523, 618, and 659; 534, 619, and 662; 535, 620, and 659; 536, 621, and 659; 537, 603, and 671; 523, 62 2, and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671; 539, 591, and 662; 523, 591, and 670; 523, 611, and 659;537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610, and 674; 523, 634, and 671; 545, 591, and 661; 523, 635 , and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616, and 660; 523, 602, and 677; 523, 616, and 659; 523, 630 , and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678; 571, 630, and 663; 572, 591, and 664; 573, 591, and 670;523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 523, 595, and 659; 523, 600, and 659; 523, 649, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 591, and 659; 584, 591, and 671; 523 , 591, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; 560, 591, and 6 The polypeptide according to claim 24, comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 59; 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or 523, 606, and 680.

26. The VLs are, respectively, sequence numbers 523, 591, and 656; 523, 592, and 657; 523, 593, and 658; 523, 591, and 659; 523, 594, and 659; 523, 595, and 660; 523, 596, and 659; 524, 591, and 659; 523, 591, and 661; 523, 591, and 662; 523, 597, and 658; 523, 598, and 662; 525, 599, and 659; 523, 600, and 663; 523, 591, and 664; 523, 601 , and 659; 526, 597, and 665; 523, 602, and 659; 527, 591, and 661; 528, 591, and 662; 529, 603, and 662; 523, 604, and 666; 523, 605, and 667; 530, 606, and 659; 523, 607, and 659; 523, 608, and 659; 523, 609, and 662; 531, 591, and 663; 523, 608, and 661; 523, 610, and 659; 523, 611, and 668; 523, 611, and 658; 523, 596, and 662; 523, 612, and 667; 523, 613, and 668; 523, 591, and 669; 523, 591, and 663; 523, 614, and 659; 532, 615, and 662; 523, 616, and 670; 523, 591, and 668; 523, 617, and 671; 533, 595, and 659; 523, 618, and 659; 534, 619, and 662; 535, 620, and 659; 536, 621, and 659; 537, 603, and 671; 523, 622 , and 659; 523, 617, and 659; 523, 623, and 659; 538, 607, and 672; 523, 616, and 671; 523, 624, and 659; 523, 620, and 659; 523, 625, and 660; 523, 602, and 671; 523, 611, and 663; 523, 626, and 670; 523, 606, and 673; 523, 627, and 671; 523, 628, and 671; 539, 591, and 662; 523, 591, and 670; 523, 611, and 659;537, 613, and 671; 540, 598, and 659; 541, 591, and 659; 542, 629, and 659; 523, 591, and 671; 523, 630, and 667; 543, 631, and 659; 523, 632, and 664; 523, 597, and 661; 523, 633, and 659; 544, 607, and 659; 523, 630, and 659; 523, 591, and 665; 523, 610, and 674; 523, 634, and 671; 545, 591, and 661; 523, 635 , and 667; 546, 591, and 659; 523, 636, and 664; 547, 591, and 663; 548, 619, and 659; 523, 637, and 659; 523, 638, and 659; 549, 591, and 659; 550, 591, and 675; 551, 591, and 659; 552, 591, and 659; 553, 591, and 676; 554, 591, and 659; 523, 591, and 676; 555, 606, and 659; 556, 591, and 659; 523, 637, and 677; 557, 591, and 659; 558, 591, and 659; 523, 591, and 677; 553, 639, and 659; 549, 591, and 667; 559, 617, and 666; 523, 591, and 672; 523, 606, and 659; 523, 592, and 670; 560, 591, and 667; 530, 591, and 659; 561, 603, and 666; 562, 591, and 659; 523, 616, and 660; 523, 602, and 677; 523, 616, and 659; 523, 630 , and 660; 563, 591, and 659; 523, 640, and 659; 564, 591, and 659; 523, 641, and 677; 565, 591, and 662; 523, 642, and 664; 566, 643, and 659; 523, 644, and 667; 567, 606, and 659; 568, 591, and 659; 569, 645, and 659; 523, 603, and 659; 570, 628, and 678; 571, 630, and 663; 572, 591, and 664; 573, 591, and 670;523, 646, and 659; 574, 591, and 670; 523, 647, and 667; 575, 591, and 659; 537, 591, and 659; 523, 635, and 670; 576, 591, and 659; 577, 606, and 659; 523, 648, and 679; 523, 592, and 659; 523, 595, and 659; 523, 600, and 659; 523, 649, and 664; 578, 641, and 659; 523, 591, and 667; 523, 606, and 660; 523, 650, and 656; 579, 621, and 671; 580, 591, and 660; 527, 591, and 659; 581, 595, and 659; 582, 640, and 664; 583, 651, and 659; 534, 591, and 659; 584, 591, and 671; 523 , 591, and 680; 523, 591, and 681; 585, 591, and 659; 523, 649, and 659; 586, 591, and 666; 587, 652, and 659; 588, 638, and 659; 523, 623, and 664; 523, 591, and 682; 553, 653, and 659; 523, 654, and 659; 523, 648, and 659; 560, 591, and 6 The polypeptide according to claim 24, comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences described in 59; 580, 606, and 659; 589, 593, and 661; 523, 637, and 676; 590, 622, and 659; 555, 591, and 661; 523, 615, and 672; 523, 626, and 668; 523, 655, and 659; or 523, 606, and 680.

27. The polypeptide according to claim 24, wherein the VL comprises any one of the amino acid sequences described in SEQ ID NOs: 181 to 368.

28. The antibody or polypeptide according to any one of the prior claims, wherein the antibody or polypeptide is conjugated with a cytotoxic agent, a cell proliferation inhibitor, a toxin, a radionuclide, or a detectable label.

29. A polynucleotide encoding the VH and / or VL of an antibody according to any one of claims 1 to 19, or a polypeptide according to any one of claims 20 to 27.

30. A vector comprising the polynucleotide described in claim 29.

31. The vector according to claim 30, wherein the vector is an adeno-associated virus (AAV) vector.

32. Recombinant host cells, (a) The polynucleotide according to claim 29, (b) The vector according to claim 30 or 31, (c) A first polynucleotide encoding the VH or heavy chain of the antibody according to any one of claims 1 to 19, and a second polynucleotide encoding the VL or light chain of the antibody according to any one of claims 1 to 19, or (d) Recombinant host cell comprising a first vector comprising a first polynucleotide encoding the VH or heavy chain of the antibody according to any one of claims 1 to 19, and a second vector comprising a second polynucleotide encoding the VL or light chain of the antibody according to any one of claims 1 to 19.

33. A pharmaceutical composition comprising an antibody or polypeptide according to any one of claims 1 to 28, a polynucleotide according to claim 29, a vector according to claim 30 or 31, or a host cell according to claim 32, and a pharmaceutically acceptable carrier or excipient.

34. A method for producing an antibody or polypeptide, the method comprising culturing the host cell described in claim 32 under preferred conditions such that a polynucleotide is expressed and the antibody or polypeptide is produced.

35. A method for inhibiting the activity of PC in a subject, the method comprising administering to the subject an effective amount of an antibody or polypeptide according to any one of claims 1 to 28, a polynucleotide according to claim 29, a vector according to claim 30 or 31, a host cell according to claim 32, or a pharmaceutical composition according to claim 33.

36. A method for treating an inflammatory disorder or degenerative disease in a subject, the method comprising administering to the subject an effective amount of an antibody or polypeptide according to any one of claims 1 to 28, a polynucleotide according to claim 29, a vector according to claim 30 or 31, a host cell according to claim 32, or a pharmaceutical composition according to claim 33.

37. The method according to claim 36, wherein the inflammatory disorder or degenerative disease is selected from the group consisting of myocardial infarction-induced reperfusion injury, Kawasaki disease, non-alcoholic steatohepatitis (NASH), organ transplant rejection, atherosclerosis, type 1 or type 2 diabetes mellitus, rheumatoid arthritis, osteoporosis, acute lung injury, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pain, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), age-related macular degeneration (AMD), stroke, Huntington's disease, frontotemporal dementia (FTD), multiple sclerosis (MS), isolated demyelinating disease of the central nervous system, osteoarthritis, Crohn's disease, and organ reperfusion injury such as ulcerative colitis.

38. Use of an antibody or polypeptide according to any one of claims 1 to 28, a polynucleotide according to claim 29, a vector according to claim 30 or 31, a host cell according to claim 32, or a pharmaceutical composition according to claim 33 for the manufacture of a pharmaceutical for treating an inflammatory disorder or a degenerative disease in a subject requiring treatment of an inflammatory disorder or a degenerative disease.

39. The use according to claim 38, wherein the inflammatory disorder or degenerative disease is selected from the group consisting of myocardial infarction-induced reperfusion injury, Kawasaki disease, non-alcoholic steatohepatitis (NASH), organ transplant rejection, atherosclerosis, type 1 or type 2 diabetes mellitus, rheumatoid arthritis, osteoporosis, acute lung injury, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pain, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), age-related macular degeneration (AMD), stroke, Huntington's disease, frontotemporal dementia (FTD), multiple sclerosis (MS), isolated demyelinating disease of the central nervous system, osteoarthritis, Crohn's disease, and organ reperfusion injury such as ulcerative colitis.

40. An antibody or polypeptide according to any one of claims 1 to 28, a polynucleotide according to claim 29, a vector according to claim 30 or 31, a host cell according to claim 32, or a pharmaceutical composition according to claim 33, for use in medical applications.

41. An antibody or polypeptide according to any one of claims 1 to 28, a polynucleotide according to claim 29, a vector according to claim 30 or 31, a host cell according to claim 32, or a pharmaceutical composition according to claim 33, for use in the treatment of an inflammatory disorder or degenerative disease in a subject requiring treatment for an inflammatory disorder or degenerative disease.

42. The use according to claim 41, wherein the inflammatory disorder or degenerative disease is selected from the group consisting of myocardial infarction-induced reperfusion injury, Kawasaki disease, non-alcoholic steatohepatitis (NASH), organ transplant rejection, atherosclerosis, type 1 or type 2 diabetes mellitus, rheumatoid arthritis, osteoporosis, acute lung injury, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pain, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), age-related macular degeneration (AMD), stroke, Huntington's disease, frontotemporal dementia (FTD), multiple sclerosis (MS), isolated demyelinating disease of the central nervous system, osteoarthritis, Crohn's disease, and organ reperfusion injury such as ulcerative colitis.

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