Identification and engineering of antibodies with variant Fc regions and methods of using same
First Claim
1. An polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said polypeptide binds an Fcγ
- R with an altered affinity relative to a polypeptide comprising a wild-type Fc region, provided that said at least one amino acid modification is not solely a substitution at any of positions 255, 258, 267, 269, 270, 276, 278, 280, 283, 285, 289, 292, 293, 294, 295, 296, 300, 303, 305, 307, 309, 322, 329, 332, 331, 337, 338, 340, 373, 376, 416, 419, 434, 435, 437, 438, 439 or does not have an alanine at any of positions 256, 290, 298, 312, 333, 334, 359, 360, 326, or 430;
a lysine at position 330;
a threonine at position 339;
a methionine at position 320;
a serine at position 326;
an asparagine at position 326;
an aspartic acid at position 326;
a glutamic acid at position 326;
a glutamine at position 334;
a glutamic acid at position 334;
a methionine at position 334;
a histidine at position 334;
a valine at position 334;
or a leucine at position 334;
a lysine at position 335 an asparagine at position 268;
a glutamine at position 272;
a glutamine, serine, or aspartic acid at position 286;
a serine at position 290;
a methionine, glutamine, glutamic acid, or arginine at position 320;
a glutamic acid at position 322;
a serine, glutamic acid, or aspartic acid at position 326;
a lysine at position 330;
a glutamine at position 335;
or a methionine at position 301.
3 Assignments
0 Petitions
Accused Products
Abstract
The present invention relates to molecules, particularly polypeptides, more particularly immunoglobulins (e.g., antibodies), comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, which variant Fc region binds FcγRIIA and/or FcγRIIA with a greater affinity, relative to a comparable molecule comprising the wild-type Fc region. The molecules of the invention are particularly useful in preventing, treating, or ameliorating one or more symptoms associated with a disease, disorder, or infection. The molecules of the invention are particularly useful for the treatment or prevention of a disease or disorder where an enhanced efficacy of effector cell function (e.g., ADCC) mediated by FcγR is desired, e.g., cancer, infectious disease, and in enhancing the therapeutic efficacy of therapeutic antibodies the effect of which is mediated by ADCC.
335 Citations
83 Claims
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1. An polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said polypeptide binds an Fcγ
- R with an altered affinity relative to a polypeptide comprising a wild-type Fc region, provided that said at least one amino acid modification is not solely a substitution at any of positions 255, 258, 267, 269, 270, 276, 278, 280, 283, 285, 289, 292, 293, 294, 295, 296, 300, 303, 305, 307, 309, 322, 329, 332, 331, 337, 338, 340, 373, 376, 416, 419, 434, 435, 437, 438, 439 or does not have an alanine at any of positions 256, 290, 298, 312, 333, 334, 359, 360, 326, or 430;
a lysine at position 330;
a threonine at position 339;
a methionine at position 320;
a serine at position 326;
an asparagine at position 326;
an aspartic acid at position 326;
a glutamic acid at position 326;
a glutamine at position 334;
a glutamic acid at position 334;
a methionine at position 334;
a histidine at position 334;
a valine at position 334;
or a leucine at position 334;
a lysine at position 335 an asparagine at position 268;
a glutamine at position 272;
a glutamine, serine, or aspartic acid at position 286;
a serine at position 290;
a methionine, glutamine, glutamic acid, or arginine at position 320;
a glutamic acid at position 322;
a serine, glutamic acid, or aspartic acid at position 326;
a lysine at position 330;
a glutamine at position 335;
or a methionine at position 301. - View Dependent Claims (5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 23, 24, 25, 37, 38, 42, 43, 44, 45, 46, 56, 57, 67, 68, 69, 70, 71, 72, 73, 74)
- R with an altered affinity relative to a polypeptide comprising a wild-type Fc region, provided that said at least one amino acid modification is not solely a substitution at any of positions 255, 258, 267, 269, 270, 276, 278, 280, 283, 285, 289, 292, 293, 294, 295, 296, 300, 303, 305, 307, 309, 322, 329, 332, 331, 337, 338, 340, 373, 376, 416, 419, 434, 435, 437, 438, 439 or does not have an alanine at any of positions 256, 290, 298, 312, 333, 334, 359, 360, 326, or 430;
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2. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said polypeptide specifically binds Fcγ
- RIIIA with a greater affinity than a comparable polypeptide comprising the wild-type Fc region binds Fcγ
RIIIA, provided that said at least one amino acid modification is not solely a substitution at positions 329, 331, or 332, or does not have an alanine at any of positions 256, 290, 298, 312, 326, 333, 334, 359, 360, or 430;
a lysine at position 330;
a threonine at position 339;
a methionine at position 320;
a serine at position 326;
an asparagine at position 326;
an aspartic acid at position 326;
a glutamic acid at position 326;
a glutamine at position 334;
a glutamic acid at position 334;
a methionine at position 334;
a histidine at position 334;
a valine at position 334;
a lysine at position 330;
a lysine at position 335;
or a leucine at position 334. - View Dependent Claims (3)
- RIIIA with a greater affinity than a comparable polypeptide comprising the wild-type Fc region binds Fcγ
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4. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said polypeptide specifically binds Fcγ
- RIIIA with a greater affinity than a comparable polypeptide comprising the wild-type Fc region binds Fcγ
RIIIA, and said polypeptide further specifically binds Fcγ
RIIB with a lower affinity than a comparable polypeptide comprising the wild-type Fc region binds Fcγ
RIIB, provided that said variant Fc region does not have an alanine at any of positions 256, 298, 333, or 334.
- RIIIA with a greater affinity than a comparable polypeptide comprising the wild-type Fc region binds Fcγ
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18. An antibody comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said antibody specifically binds Fcγ
- RIIIA with a greater affinity than a comparable antibody comprising the wild-type Fc region binds Fcγ
RIIIA, and said antibody further specifically binds Fcγ
RIIB with a lower affinity than a comparable antibody comprising the wild-type Fc region binds Fcγ
RIIB, provided that said variant Fc region does not have an alanine at any of positions 256, 298, 333, or 334.
- RIIIA with a greater affinity than a comparable antibody comprising the wild-type Fc region binds Fcγ
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26. A therapeutic antibody that specifically binds a cancer antigen, said therapeutic antibody comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said therapeutic antibody specifically binds Fcγ
- RIIIA with a greater affinity than the therapeutic antibody comprising the wild-type Fc region binds Fcγ
RIIIA, provided that said at least one amino acid modification is not solely a substitution at positions 329, 331, or 332, or does not have an alanine at any of positions 256, 290, 298, 312, 333, 334, 359, 360, or 430;
a lysine at position 330;
a threonine at position 339;
a methionine at position 320;
a serine at position 326;
an asparagine at position 326;
an aspartic acid at position 326;
a glutamic acid at position 326;
a glutamine at position 334;
a glutamic acid at position 334;
a methionine at position 334;
a histidine at position 334;
a valine at position 334;
or a leucine at position 334. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 39, 40, 41)
- RIIIA with a greater affinity than the therapeutic antibody comprising the wild-type Fc region binds Fcγ
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27. A therapeutic antibody that specifically binds a cancer antigen, said therapeutic antibody comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said therapeutic antibody specifically binds Fcγ
- RIIIA with a greater affinity than a therapeutic antibody comprising the wild-type Fc region binds Fcγ
RIIIA, and said therapeutic antibody further specifically binds Fcγ
RIIB with a lower affinity than a therapeutic antibody comprising the wild-type Fc region binds Fcγ
RIIB, provided that said variant Fc region does not solely have an alanine at any of positions 256, 298, 333, or 334.
- RIIIA with a greater affinity than a therapeutic antibody comprising the wild-type Fc region binds Fcγ
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47. A method for producing a tetrameric Fcγ
- R complex, wherein said tetrameric complex has an enhanced affinity for an Fc region, relative to the affinity of a monomeric Fcγ
R for the Fc region, comprising;
(a) producing a fusion protein, such that a 15 amino acid AVITAG sequence is operably linked to the soluble region of Fcγ
R;
(b) biotinylating the protein produced in step (a);
(c) mixing the biotinylated protein produced in step (b) with streptaividn-phycoerythin in an appropriate molar ratio, such that a tetrameric Fcγ
R complex is formed. - View Dependent Claims (48, 49, 50, 51, 52, 53, 54, 55)
- R complex, wherein said tetrameric complex has an enhanced affinity for an Fc region, relative to the affinity of a monomeric Fcγ
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58. A method of treating an autoimmune disorder in a patient in need thereof, said method comprising administering to said patient a therapeutically effective amount of a molecule comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild type Fc region, such that said molecule specifically binds Fcγ
- RIIB with a greater affinity than a comparable molecule comprising the wild type Fc region, and said molecule further specifically binds Fcγ
RIIIA with a lower affinity than a comparable molecule comprising the wild type Fc region. - View Dependent Claims (59, 60, 61, 62, 63, 64, 65)
- RIIB with a greater affinity than a comparable molecule comprising the wild type Fc region, and said molecule further specifically binds Fcγ
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66. A method of treating an infectious disease in a patient in need thereof, said method comprising administering to said patient a therapeutically effective amount of a molecule comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild type Fc region, such that said molecule specifically binds Fcγ
- RIIB with a greater affinity than a comparable molecule comprising the wild type Fc region, and said molecule further specifically binds Fcγ
RIIIA with a lower affinity than a comparable molecule comprising the wild type Fc region.
- RIIB with a greater affinity than a comparable molecule comprising the wild type Fc region, and said molecule further specifically binds Fcγ
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75. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, such that said polypeptide specifically binds Fcγ
- RIIIA with a greater affinity than a comparable polypeptide comprising the wild-type Fc region, and said polypeptide further specifically binds Fcγ
RIIB with a lower affinity than a comparable polypeptide comprising the wild type Fc region binds Fcγ
RIIB, wherein said at least one amino acid modification comprises a set of substitutions selected from the group consisting of a substitution at position 243 with isoleucine and at position 379 with leucine;
a substitution at position 288 with asparagine, at position 330 with serine and at position 396 with leucine;
a substitution at position 243 with leucine and at position 255 with leucine;
a substitution at position 288 with methionine and at position 334 with glutamic acid;
a substitution at position 316 with aspartic acid, at position 378 with valine, and at position 399 with glutamic acid;
a substitution at position 315 with isoleucine, at position 379 with methionine, and at position 399 with glutamic acid;
a substitution at position 243 with isoleucine, at position 379 with leucine, and at position 420 with valine;
a substitution at position 392 with threonine and at position 396 with leucine;
a substitution at position 293 with valine, at position 295 with glutamic acid, and at position 327 with threonine;
a substitution at position 268 with asparagine and at position 396 with leucine;
a substitution at position 319 with phenylalanine, at position 352 with leucine, and at position 396 with leucine;
a substitution at position 248 with methionine;
a substitution at position 247 with leucine and at position 420 with valine;
a substitution 334 with glutamic acid and at position 292 with leucine. - View Dependent Claims (76, 77)
- RIIIA with a greater affinity than a comparable polypeptide comprising the wild-type Fc region, and said polypeptide further specifically binds Fcγ
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78. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification, wherein said at least one amino acid modification comprises substitution at position 255 with leucine and at position 396 with leucine.
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79. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification, wherein said at least one amino acid modification comprises substitution at position 370 with glutamic acid and at position 396 with leucine.
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80. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification, wherein said at least one amino acid modification comprises substitution at position 392 with threonine and at position 396 with leucine.
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81. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification, wherein said at least one amino acid modification comprises substitution at position 221 with glutamic acid, at position 270 with glutamic acid, at position 308 with alanine, at position 311 with histidine, at position 396 with leucine, and at position 402 with aspartic acid.
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82. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification, wherein said at least one amino acid modification comprises substitution at position 243 with leucine, at position 305 with isoleucine, at position 378 with aspartic acid, at position 404 with serine, and at position 396 with leucine.
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83. A polypeptide comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification, wherein said at least one amino acid modification comprises substitution at position 284 with methionine, at position 298 with asparagine, at position 334 with glutamic acid, at position 355 with tryptophan, and at position 416 with threonine.
Specification