Drug containing carrier into cell for forming immune complex

Abstract

The present inventors discovered that by forming a large immune complex comprising antigens containing two or more antigenic binding units (epitopes) and two or more antigen-binding molecules (for example, antibodies), elimination from the plasma of the antigens containing two or more antigenic binding units can be accelerated. Moreover, they found that by using this characteristic and by further using antigen-binding molecules having an ion-dependent antigen-binding activity, elimination of the antigens can further be accelerated and the above problem can be solved.

1 - 29 . (canceled) 
     
     
         30 . A method of reducing the amount of an antigen in the plasma of a subject, the method comprising
 identifying a subject as being in need of a reduction in the amount of the antigen in the subject's plasma; and   administering to the subject's plasma an antigen-binding molecule comprising (i) an Fc region, and (ii) two or more antigen-binding domains, including a first antigen-binding domain that binds to a first epitope on the antigen and a second antigen-binding domain that binds to a second epitope on the antigen, wherein   (a) the first and second epitopes may be identical or different,   (b) the antigen-binding activity of at least one of the antigen-binding domains varies depending on pH or on calcium ion concentration, and   (c) the antigen-binding molecule forms an immune complex comprising two or more copies of the antigen-binding molecule and two or more copies of the antigen.   
     
     
         31 . The method of  claim 30 , wherein the antigen-binding activity of at least one of the antigen-binding domains varies depending on calcium ion concentration. 
     
     
         32 . The method of  claim 31 , wherein the antigen-binding activity of at least one of the antigen-binding domains is lower at a calcium ion concentration between 0.5 μM and 10 μM than at a calcium ion concentration between 200 μM and 5 mM. 
     
     
         33 . The method of  claim 30 , wherein the antigen-binding activity of at least one of the antigen-binding domains varies with pH. 
     
     
         34 . The method of  claim 33 , wherein the antigen-binding activity of at least one of the antigen-binding domains is lower at a pH in the range of 4.0 to 6.5 than at a pH in the range of 6.7 to 10.0. 
     
     
         35 . The method of  claim 30 , wherein the antigen is a multimer comprising two or more subunits that may be the same or different. 
     
     
         36 . The method of  claim 35 , wherein each of at least two of the subunits comprises at least one of the epitopes. 
     
     
         37 . The method of  claim 35 , wherein the antigen is any one of GDF, GDF-1, GDF-3 (Vgr-2), GDF-5 (BMP-14, CDMP-1), GDF-6 (BMP-13, CDMP-2), GDF-7 (BMP-12, CDMP-3), GDF-8 (myostatin), GDF-9, GDF-15 (MIC-1), TNF, TNF-alphabeta, TNF-beta2, TNFSF10 (TRAIL, Apo-2 ligand, TL2), TNFSF11 (TRANCE/RANK ligand, ODF, OPG ligand), TNFSF12 (TWEAK, Apo-3 ligand, DR3 ligand), TNFSF13 (APRIL, TALL2), TNFSF13B (BAFF, BLYS, TALL1, THANK, TNFSF20), TNFSF14 (LIGHT, HVEM ligand, LTg), TNFSF15 (TL1A/VEGI), TNFSF18 (GITR ligand, AITR ligand, TL6), TNFSF1A (TNF-a, Cachectin, DIF, TNFSF2), TNFSF1B (TNF-b, LTa, TNFSF1), TNFSF3 (LTb, TNFC, p33), TNFSF4 (OX40 ligand, gp34, TXGP1), TNFSF5 (CD40 ligand, CD154, gp39, HIGM1, IMD3, TRAP), TNFSF6 (Fas ligand, Apo-1 ligand, APT1 ligand), TNFSF7 (CD27 ligand, CD70), TNFSF8 (CD30 ligand, CD153), TNFSF9 (4-1BB ligand, CD137 ligand), VEGF, IgE, IgA, IgG, IgM, RANKL, TGF-alpha, TGF-beta, TGF-beta Pan Specific, or IL-8. 
     
     
         38 . The method of  claim 30 , wherein the antigen is a monomer comprising the first and second epitopes. 
     
     
         39 . The method of  claim 30 , wherein the antigen-binding molecule is a multispecific or multiparatopic antigen-binding molecule. 
     
     
         40 . The method of  claim 30 , wherein the Fc region of (i) comprises the sequence of any one of SEQ ID NOs: 13, 14, 15, and 16. 
     
     
         41 . The method of  claim 30 , wherein the Fc region of (i) has, at a pH between 4 and 6.5, an FcRn-binding activity greater than that of a second Fc region at the same pH, wherein the second Fc region is represented by any one of SEQ ID NOs: 13, 14, 15, and 16. 
     
     
         42 . The method of  claim 41 , wherein the Fc region of (i) comprises the amino acid sequence of any one of SEQ ID NOs: 13, 14, 15, and 16, except with an amino acid substitution at one or more positions selected from positions 238, 244, 245, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 260, 262, 265, 270, 272, 279, 283, 285, 286, 288, 293, 303, 305, 307, 308, 309, 311, 312, 314, 316, 317, 318, 332, 339, 340, 341, 343, 356, 360, 362, 375, 376, 377, 378, 380, 382, 385, 386, 387, 388, 389, 400, 413, 415, 423, 424, 427, 428, 430, 431, 433, 434, 435, 436, 438, 439, 440, 442, and 447 (EU numbering). 
     
     
         43 . The method of  claim 42 , wherein the Fc region of (i) comprises, at one or more of the following positions, the indicated amino acid (all positions by EU numbering):
 Leu at position 238;   Leu at position 244;   Arg at position 245;   Pro at position 249;   Gln or Glu at position 250;   Arg, Asp, Glu, or Leu at position 251;   Phe, Ser, Thr, or Tyr at position 252;   Ser or Thr at position 254;   Arg, Gly, Ile, or Leu at position 255;   Ala, Arg, Asn, Asp, Gln, Glu, Pro, or Thr at position 256;   Ala, Ile, Met, Asn, Ser, or Val at position 257;   Asp at position 258;   Ser at position 260;   Leu at position 262;   Lys at position 270;   Leu or Arg at position 272;   Ala, Asp, Gly, His, Met, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr at position 279;   Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr at position 283;   Asn at position 285;   Phe at position 286;   Asn or Pro at position 288;   Val at position 293;   Ala, Glu, Gln, or Met at position 307;   Ile, Pro, or Thr at position 308;   Ala, Glu, Ile, Lys, Leu, Met, Ser, Val, or Trp at position 311;   Pro at position 309;   Ala, Asp, or Pro at position 312;   Ala or Leu at position 314;   Lys at position 316;   Pro at position 317;   Asn or Thr at position 318;   Phe, His, Lys, Leu, Met, Arg, Ser, or Trp at position 332;   Asn, Thr, or Trp at position 339;   Pro at position 341;   Glu, His, Lys, Gln, Arg, Thr, or Tyr at position 343;   Arg at position 375;   Gly, Ile, Met, Pro, Thr, or Val at position 376;   Lys at position 377;   Asp, Asn, or Val at position 378;   Ala, Asn, Ser, or Thr at position 380;   Phe, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 382;   Ala, Arg, Asp, Gly, His, Lys, Ser, or Thr at position 385;   Arg, Asp, Ile, Lys, Met, Pro, Ser, or Thr at position 386;   Ala, Arg, His, Pro, Ser, or Thr at position 387;   Asn, Pro, or Ser at position 389;   Asn at position 423;   Asn at position 427;   Leu, Met, Phe, Ser, or Thr at position 428;   Ala, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr at position 430;   His or Asn at position 431;   Arg, Gln, His, Ile, Lys, Pro, or Ser at position 433;   Ala, Gly, His, Phe, Ser, Trp, or Tyr at position 434;   Arg, Asn, His, Ile, Leu, Lys, Met, or Thr at position 436;   Lys, Leu, Thr, or Trp at position 438;   Lys at position 440; and   Lys at position 442.   
     
     
         44 . The method of  claim 30 , wherein the Fc region of (i) has, at a pH between 6.7 and 10.0, an FcRn-binding activity greater than that of a second Fc region at the same pH, wherein the second Fc region is represented by any one of SEQ ID NOs: 13, 14, 15, and 16. 
     
     
         45 . The method of  claim 44 , wherein the Fc region of (i) comprises the amino acid sequence of any one of SEQ ID NOs: 13, 14, 15, and 16, except with an amino acid substitution at one or more positions selected from positions 237, 248, 250, 252, 254, 255, 256, 257, 258, 265, 286, 289, 297, 298, 303, 305, 307, 308, 309, 311, 312, 314, 315, 317, 332, 334, 360, 376, 380, 382, 384, 385, 386, 387, 389, 424, 428, 433, 434, and 436 (EU numbering). 
     
     
         46 . The method of  claim 45 , wherein the Fc region of (i) comprises, at one or more of the following positions, the indicated amino acid (all positions by EU numbering):
 Met at position 237;   Ile at position 248;   Ala, Phe, Ile, Met, Gln, Ser, Val, Trp, or Tyr at position 250;   Phe, Trp, or Tyr at position 252;   Thr at position 254;   Glu at position 255;   Asp, Asn, Glu, or Gln at position 256;   Ala, Gly, Ile, Leu, Met, Asn, Ser, Thr, or Val at position 257;   His at position 258;   Ala at position 265;   Ala or Glu at position 286;   His at position 289;   Ala at position 297;   Ala at position 303;   Ala at position 305;   Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp, or Tyr at position 307;   Ala, Phe, Ile, Leu, Met, Pro, Gln, or Thr at position 308;   Ala, Asp, Glu, Pro, or Arg at position 309;   Ala, His, or Ile at position 311;   Ala or His at position 312;   Lys or Arg at position 314;   Ala, Asp, or His at position 315;   Ala at position 317;   Val at position 332;   Leu at position 334;   His at position 360;   Ala at position 376;   Ala at position 380;   Ala at position 382;   Ala at position 384;   Asp or His at position 385;   Pro at position 386;   Glu at position 387;   Ala or Ser at position 389;   Ala at position 424;   Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Ser, Thr, Val, Trp, or Tyr at position 428;   Lys at position 433;   Ala, Phe, His, Ser, Trp, or Tyr at position 434;   His, Ile, Leu, Phe, Thr, or Val at position 436.   
     
     
         47 . The method of  claim 30 , wherein the Fc region of (i) has a higher Fcγ receptor-binding activity than does the Fc region of a native human IgG. 
     
     
         48 . The method of  claim 47 , wherein the amino acid sequence of the Fc region of (i) differs from the amino acid sequence of the Fc region of the native human IgG at one or more positions, including at least one of the following positions (EU numbering): 221, 222, 223, 224, 225, 227, 228, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 251, 254, 255, 256, 258, 260, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 288, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 311, 313, 315, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 339, 376, 377, 378, 379, 380, 382, 385, 392, 396, 421, 427, 428, 429, 434, 436, 440. 
     
     
         49 . The method of  claim 48 , wherein the Fc region of (i) comprises, at one or more of the following positions, the indicated amino acid (all positions by EU numbering):
 Lys or Tyr at position 221;   Phe, Trp, Glu, or Tyr at position 222;   Phe, Trp, Glu, or Lys at position 223;   Phe, Trp, Glu, or Tyr at position 224;   Glu, Lys, or Trp at position 225;   Glu, Gly, Lys, or Tyr at position 227;   Glu, Gly, Lys, or Tyr at position 228;   Ala, Glu, Gly, or Tyr at position 230;   Glu, Gly, Lys, Pro, or Tyr at position 231;   Glu, Gly, Lys, or Tyr at position 232;   Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 233;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Tip, or Tyr at position 234;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Tip, or Tyr at position 235;   Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 236;   Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 237;   Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 238;   Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr at position 239;   Ala, Ile, Met, or Thr at position 240;   Asp, Glu, Leu, Arg, Trp, or Tyr at position 241;   Leu, Glu, Leu, Gln, Arg, Trp, or Tyr at position 243;   His at position 244;   Ala at position 245;   Asp, Glu, His, or Tyr at position 246;   Ala, Phe, Gly, His, Ile, Leu, Met, Thr, Val, or Tyr at position 247;   Glu, His, Gln, or Tyr at position 249;   Glu or Gln at position 250;   Phe at position 251;   Phe, Met, or Tyr at position 254;   Glu, Leu, or Tyr at position 255;   Ala, Met, or Pro at position 256;   Asp, Glu, His, Ser, or Tyr at position 258;   Asp, Glu, His, or Tyr at position 260;   Ala, Glu, Phe, Ile, or Thr at position 262;   Ala, Ile, Met, or Thr at position 263;   Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr at position 264;   Ala, Leu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 265;   Ala, Ile, Met, or Thr at position 266;   Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr at position 267;   Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Pro, Gln, Arg, Thr, Val, or Trp at position 268;   Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 269;   Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr at position 270;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 271;   Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 272;   Phe or Ile at position 273;   Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 274;   Leu or Trp at position 275;   Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 276;   Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or Trp at position 278;   Ala at position 279;   Ala, Gly, His, Lys, Leu, Pro, Gln, Trp, or Tyr at position 280;   Asp, Lys, Pro, or Tyr at position 281;   Glu, Gly, Lys, Pro, or Tyr at position 282;   Ala, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, or Tyr at position 283;   Asp, Glu, Leu, Asn, Thr, or Tyr at position 284;   Asp, Glu, Lys, Gln, Trp, or Tyr at position 285;   Glu, Gly, Pro, or Tyr at position 286;   Asn, Asp, Glu, or Tyr at position 288;   Asp, Gly, His, Leu, Asn, Ser, Thr, Trp, or Tyr at position 290;   Asp, Glu, Gly, His, Ile, Gln, or Thr at position 291;   Ala, Asp, Glu, Pro, Thr, or Tyr at position 292;   Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 293;   Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 294;   Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 295;   Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val at position 296;   Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 297;   Ala, Asp, Glu, Phe, His, Ile, Lys, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr at position 298;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp, or Tyr at position 299;   Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or Trp at position 300;   Asp, Glu, His, or Tyr at position 301;   Ile at position 302;   Asp, Gly, or Tyr at position 303;   Asp, His, Leu, Asn, or Thr at position 304;   Glu, Ile, Thr, or Tyr at position 305;   Ala, Asp, Asn, Thr, Val, or Tyr at position 311;   Phe at position 313;   Leu at position 315;   Glu or Gln at position 317;   His, Leu, Asn, Pro, Gln, Arg, Thr, Val, or Tyr at position 318;   Asp, Phe, Gly, His, Ile, Leu, Asn, Pro, Ser, Thr, Val, Trp, or Tyr at position 320;   Ala, Asp, Phe, Gly, His, Ile, Pro, Ser, Thr, Val, Trp, or Tyr at position 322;   Ile at position 323;   Asp, Phe, Gly, His, Ile, Leu, Met, Pro, Arg, Thr, Val, Trp, or Tyr at position 324;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 325;   Ala, Asp, Glu, Gly, Ile, Leu, Met, Asn, Pro, Gln, Ser, Thr, Val, Trp, or Tyr at position 326;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Thr, Val, Trp, or Tyr at position 327;   Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 328;   Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 329;   Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr at position 330;   Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Thr, Val, Trp, or Tyr at position 331;   Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at position 332;   Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Ser, Thr, Val, or Tyr at position 333;   Ala, Glu, Phe, Ile, Leu, Pro, or Thr at position 334;   Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Val, Trp, or Tyr at position 335;   Glu, Lys, or Tyr at position 336;   Glu, His, or Asn at position 337;   Asp, Phe, Gly, Ile, Lys, Met, Asn, Gln, Arg, Ser, or Thr at position 339;   Ala or Val at position 376;   Gly or Lys at position 377;   Asp at position 378;   Asn at position 379;   Ala, Asn, or Ser at position 380;   Ala or Ile at position 382;   Glu at position 385;   Thr at position 392;   Leu at position 396;   Lys at position 421;   Asn at position 427;   Phe or Leu at position 428;   Met at position 429;   Trp at position 434;   Ile at position 436;   Gly, His, Ile, Leu, or Tyr at position 440.   
     
     
         50 . The method of  claim 30 , wherein the Fc region has a higher binding activity toward an inhibitory Fcγ receptor than toward an activating Fcγ receptor. 
     
     
         51 . The method of  claim 50 , wherein the inhibitory Fcγ receptor is human FcγRIIb, and the activating Fcγ receptor is human FcγRIa, human FcγRIIa (R), human FcγRIIa (H), human FcγRIIIa (V), or human FcγRIIIa (F). 
     
     
         52 . The method of  claim 50 , wherein the amino acid at position 238 or 328 (EU numbering) in the Fc region of (i) is different from the amino acid in the corresponding position of a native human IgG Fc region. 
     
     
         53 . The method of  claim 52 , wherein the amino acid at position 238 of the Fc region of (i) is Asp, or the amino acid at position 328 of the Fc region of (i) is Glu (all positions by EU numbering). 
     
     
         54 . The method of  claim 53 , wherein the Fc region of (i) comprises, at one or more of the following positions, the indicated amino acid (all positions by EU numbering):
 Asp at position 233;   Tip or Tyr at position 234;   Ala, Asp, Glu, Leu, Met, Phe, Trp, or Tyr at position 237;   Asp at position 239;   Ala, Gln, or Val at position 267;   Asn, Asp, or Glu at position 268;   Gly at position 271;   Ala, Asn, Asp, Gln, Glu, Leu, Met, Ser, or Thr at position 326;   Arg, Lys, or Met at position 330;   Ile, Leu, or Met at position 323;   Asp at position 296.   
     
     
         55 . A method of reducing the amount of an antigen in the plasma of a subject, the method comprising
 identifying a subject as being in need of a reduction in the amount of the antigen in the subject's plasma; and   administering to the subject at least two different antigen-binding molecules including (i) a first antigen-binding molecule that comprises a first Fc region and a first antigen-binding domain that binds to a first epitope on the antigen, and (ii) a second antigen-binding molecule that comprises a second Fc region and a second antigen binding domain that binds to a second epitope on the antigen, wherein   (a) the first and second epitopes may be identical or different,   (b) the antigen-binding activity of at least one of the antigen-binding domains varies depending on pH or on calcium ion concentration, and   (c) when contacted with molecules of the antigen, the first and second antigen-binding molecules form an immune complex comprising the first and second antigen-binding molecules and two or more copies of the antigen.   
     
     
         56 . The method of  claim 55 , wherein the at least two different antigen-binding molecules are administered in admixture. 
     
     
         57 . A pharmaceutical composition comprising at least two different antigen-binding molecules including (i) a first antigen-binding molecule that comprises a first Fc region and a first antigen-binding domain that binds to a first epitope on the antigen, and (ii) a second antigen-binding molecule that comprises a second Fc region and a second antigen binding domain that binds to a second epitope on the antigen, wherein
 (a) the first and second epitopes may be identical or different,   (b) the antigen-binding activity of at least one of the antigen-binding domains varies depending on pH or on calcium ion concentration, and   (c) when contacted with molecules of the antigen, the first and second antigen-binding molecules form an immune complex comprising the first and second antigen-binding molecules and two or more copies of the antigen.   
     
     
         58 . A method of screening for an antigen-binding molecule able to remove an antigen from plasma, the method comprising:
 (a) providing a composition comprising multiple copies of an antigen-binding molecule that comprises an Fc region and two or more antigen-binding domains, including a first antigen-binding domain that binds to a first epitope on the antigen and a second antigen-binding domain that binds to a second epitope on the antigen, wherein
 (i) the first and second epitopes may be identical or different, and 
 (ii) the antigen-binding activity of at least one of the antigen-binding domains varies depending on pH or on calcium ion concentration; 
   (b) contacting the composition with an antigen composition comprising multiple copies of the antigen, thereby forming immune complexes comprising the antigen and the antigen-binding molecule;   (c) determining that the immune complexes formed in (b) include immune complexes containing multiple copies of the antigen-binding molecule and multiple copies of the antigen in a single immune complex; and   (d) based on the determination of (c), identifying the antigen-binding molecule as potentially being able to remove the antigen from plasma.   
     
     
         59 . The method of  claim 58 , further comprising
 (e) testing whether the antigen-binding molecule removes the antigen from a subject's plasma in vivo.   
     
     
         60 . A method of manufacturing an antigen-binding molecule able to remove an antigen from plasma, the method comprising:
 (a) providing a cell comprising nucleic acid encoding an antigen-binding molecule comprising an Fc region and two or more antigen-binding domains, including a first antigen-binding domain that binds to a first epitope on the antigen and a second antigen-binding domain that binds to a second epitope on the antigen, wherein
 (i) the first and second epitopes may be identical or different, 
 (ii) the antigen-binding activity of at least one of the antigen-binding domains varies depending on pH or on calcium ion concentration, and 
 (iii) the antigen-binding molecule forms an immune complex comprising two or more molecules of the antigen-binding molecule and two or more molecules of the antigen; 
   (b) culturing the cell, thereby producing the antigen-binding molecule by expression of the nucleic acid; and   (c) obtaining the antigen-binding molecule from the cell culture.   
     
     
         61 . The method of  claim 60 , further comprising confirming that the antigen-binding molecule obtained in (c) is able to remove the antigen from a subject's plasma in vivo. 
     
     
         62 . A method of screening for an antigen-binding molecule able to remove an antigen from plasma, the method comprising
 (a) providing a plurality of different antigen-binding molecules that bind to the antigen, each comprising an Fc region and two or more antigen-binding domains, wherein at least one of the antigen-binding domains of each molecule of the plurality has an antigen-binding activity that varies depending on pH or on calcium ion concentration;   (b) contacting each of the plurality of different antigen-binding molecules with molecules of the antigen, thereby forming immune complexes comprising antigen-binding molecules and molecules of the antigen; and   (c) selecting an antigen-binding molecule that forms an immune complex comprising at least two copies of an antigen-binding molecule and at least two molecules of the antigen.   
     
     
         63 . The method of  claim 62 , comprising
 (d) culturing a cell containing nucleic acid encoding the antigen-binding molecule selected in (c), thereby producing the selected antigen-binding molecule.   
     
     
         64 . The method of  claim 63 , comprising
 (e) administering the antigen-binding molecule produced in (d) to a subject having plasma that comprises the antigen; and   (f) confirming that the level of antigen in the plasma of the subject decreases after the antigen-binding molecule is administered.