Method for altering plasma retention and immunogenicity of antigen-binding molecule

  • US 10,618,965 B2
  • Filed: 03/30/2012
  • Issued: 04/14/2020
  • Est. Priority Date: 02/25/2011
  • Status: Active Grant
First Claim
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1. A method for reducing immunogenicity of an antibody, the method comprising:

  • identifying a first antibody comprising (a) an Fc region with a human FcRn-binding activity at pH 7.0 that is stronger than the human FcRn-binding activity at pH 7.0 of a native human IgG1 Fc region, and (b) an antigen-binding domain whose antigen-binding activity varies with pH as described in (i) below, or with calcium ion concentration as described in (ii) below;

    (i) the antigen-binding activity is lower at pH 5.8 than at pH 7.4, wherein the ratio of the KD value for antigen-binding activity at pH 5.8 to the KD value for antigen-binding activity at pH 7.4 (KD (pH 5.8)/KD (pH 7.4)) is 2 or more when the KD (pH 5.8) and KD (pH 7.4) values are determined using a surface plasmon resonance technique in which the antibody is immobilized, the antigen serves as analyte, and the following conditions are used;

    10 mM 2-(N-morpholino)ethanesulfonic acid (MES) pH 5.8 or pH 7.4, 150 mM NaCl, 0.05% polysorbate 20, at 37°

    C.;

    (ii) the antigen-binding activity is lower at a calcium ion concentration of 3 μ

    M than at a calcium ion concentration of 2 mM, wherein the ratio of the KD value for antigen-binding activity at a calcium ion concentration of 3 μ

    M to the KD value for antigen-binding activity at a calcium ion concentration of 2 mM (KD (3 μ

    M)/ KD (2 mM)) is 2 or more when the KD (3 μ

    M) and KD (2 mM) values are determined using a surface plasmon resonance technique in which the antibody is immobilized, the antigen serves as analyte, and the following conditions are used;

    10 mM N-(2-acetamido)-2-aminoethanesulfonic acid (ACES) pH 7.4, 150 mM NaCl, 0.05% polysorbate 20, and either 2 mM CaCl2 or 3 μ

    M CaCl2, at 37°

    C.;

    producing a second antibody whose ability to form a heterocomplex with two molecules of the human FcRn and one molecule of an activating Fcγ

    receptor at pH 7.4 is reduced compared to the ability of the first antibody to form such a heterocomplex at pH 7.4, and whose ability to bind to the activating Fcγ

    receptor is decreased compared to the ability of the native human IgG1 Fc region to bind to the activating Fcγ

    receptor, the second antibody being identical to the first antibody except for one or more amino acids in the Fc region, wherein the activating Fcγ

    receptor is human Fcγ

    RIa, human Fcγ

    RIIa(R), human Fcγ

    RIIa(H), human Fcγ

    RIIIa(V), or human Fcγ

    RIIIa(F), and wherein at least one of the following positions in the Fc region of the second antibody is occupied by one of the amino acid residues listed for that position (by EU numbering);

    Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Lys, Met, Phe, Pro, Ser, Thr, or Trp at position 234;

    Ala, Asn, Asp, Gln, Glu, Gly, His, Ile, Lys, Met, Pro, Ser, Thr, Val, or Arg at position 235;

    Arg, Asn, Gln, His, Leu, Lys, Met, Phe, Pro, or Tyr at position 236;

    Ala, Asn, Asp, Gln, Glu, His, Ile, Leu, Lys, Met, Pro, Ser, Thr, Val, Tyr, or Arg at position 237;

    Ala, Asn, Gln, Glu, Gly, His, Ile, Lys, Thr, Trp, or Arg at position 238;

    Gln, His, Lys, Phe, Pro, Trp, Tyr, or Arg at position 239;

    Ala, Arg, Asn, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val at position 265;

    Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Lys, Phe, Pro, Ser, Thr, Trp, or Tyr at position 266;

    Arg, His, Lys, Phe, Pro, Trp, or Tyr at position 267;

    Ala, Arg, Asn, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val at position 269;

    Ala, Arg, Asn, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val at position 270;

    Arg, His, Phe, Ser, Thr, Trp, or Tyr at position 271;

    Arg, Asn, Asp, Gly, His, Phe, Ser, Trp, or Tyr at position 295;

    Arg, Gly, Lys, or Pro at position 296;

    Ala at position 297;

    Arg, Gly, Lys, Pro, Trp, or Tyr at position 298;

    Arg, Lys, or Pro at position 300;

    Lys or Pro at position 324;

    Ala, Arg, Gly, His, Ile, Lys, Phe, Pro, Thr, Trp, Tyr, or Val at position 325;

    Arg, Gln, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val at position 327;

    Arg, Asn, Gly, His, Lys, or Pro at position 328;

    Asn, Asp, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, Val, or Arg at position 329;

    Pro or Ser at position 330;

    Arg, Gly, or Lys at position 331;

    Arg, Lys, or Pro at position 332; and

    conducting an assay to confirm that the second antibody has decreased immunogenicity compared to the first antibody.

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