ANTIGEN BINDING POLYPEPTIDES, ANTIGEN BINDING POLYPEPTIDE COMPLEXES AND METHODS OF USE THEREOF
Disclosed are antigen binding polypeptides and antigen binding polypeptide complexes (e.g., antibodies and antigen binding fragments thereof) having certain structural features. Also disclosed are polynucleotides and vectors encoding such polypeptides and polypeptide complexes; host cells, chimeric antigen receptors (CARs), immune cells, pharmaceutical compositions and kits containing such polypeptides and polypeptide complexes; and methods of using such polypeptides and polypeptide complexes.
1 - 127 . (canceled)
128 . An antigen binding polypeptide complex comprising a first polypeptide and a second polypeptide;
wherein the first polypeptide has a structure represented by:
VL1-VL2-VH2-VH1-Fc;
VH1-VH2-VL2-VL1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc; or
VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc;
wherein the second polypeptide has a structure represented by:
VL3-VL4-VH4-VH3-Fc;
VH3-VH4-VL4-VL3-Fc;
VL3-L5-VL4-L6-VH4-L7-VH3-Fc;
VH3-L5-VH4-L6-VL4-L7-VL3-Fc;
VL3-L5-VL4-L6-VH4-L7-VH3-L8-Fc; or
VH3-L5-VH4-L6-VL4-L7-VL3-L8-Fc; and
wherein:
VL1 is a first immunoglobulin light chain variable region;
VL2 is a second immunoglobulin light chain variable region;
VL3 is a third immunoglobulin light chain variable region;
VL4 is a fourth immunoglobulin light chain variable region;
VH1 is a first immunoglobulin heavy chain variable region;
VH2 is a second immunoglobulin heavy chain variable region;
VH3 is a third immunoglobulin heavy chain variable region;
VH4 is a fourth immunoglobulin heavy chain variable region;
Fc is a region comprising an immunoglobulin heavy chain constant region 2 (CH2), an immunoglobulin heavy chain constant region 3 (CH3), and optionally, an immunoglobulin hinge; and
L1, L2, L3, L4, L5, L6, L7 and L8 are amino acid linkers.
129 . The antigen binding polypeptide complex of claim 128 ,
wherein the first polypeptide has a structure represented by:
VL1-VL2-VH2-VH1-CH1-Fc;
VH1-VH2-VL2-VL1-CH1-Fc;
VL1-VL2-VH2-VH1-CL-Fc;
VH1-VH2-VL2-VL1-CL-Fc;
VL1-VL2-VH2-VH1-CH1-CL-Fc;
VH1-VH2-VL2-VL1-CH1-CL-Fc;
VL1-VL2-VH2-VH1-CL-CH1-Fc;
VH1-VH2-VL2-VL1-CL-CH1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-L6-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-L6-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-L6-Fc; or
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-L6-Fc;
wherein the second polypeptide has a structure represented by:
VL3-VL4-VH4-VH3-CH1-Fc;
VH3-VH4-VL4-VL3-CH1-F c;
VL3-VL4-VH4-VH3-CL-Fc;
VH3-VH4-VL4-VL3-CL-F c;
VL3-VL4-VH4-VH3-CH1-CL-Fc;
VH3-VH4-VL4-VL3-CH1-CL-F c;
VL3-VL4-VH4-VH3-CL-CH1-Fc;
VH3-VH4-VL4-VL3-CL-CH1-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CH1-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CH1-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CL-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CL-Fc;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CH1-L11-CL-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CH1-L11-CL-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CL-L11-CH1-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CL-L11-CH1-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CH1-L11-CL-L12-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CH1-L11-CL-L12-Fc;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CL-L11-CH1-L12-Fc; or
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CL-L11-CH1-L12-F c; and
wherein:
CH1 is an immunoglobulin heavy chain constant region 1;
CL is an immunoglobulin light chain constant region; and
L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11 and L12 are amino acid linkers.
130 . An antigen binding polypeptide complex comprising a first polypeptide and a second polypeptide;
wherein the first polypeptide has a structure represented by:
VL1-VL2-VH2-VH1-Fc;
VH1-VH2-VL2-VL1-Fc;
VL1-VL2-VH2-VH1-CH1-Fc;
VH1-VH2-VL2-VL1-CH1-Fc;
VL1-VL2-VH2-VH1-CL-Fc;
VH1-VH2-VL2-VL1-CL-Fc;
VL1-VL2-VH2-VH1-CH1-CL-Fc;
VH1-VH2-VL2-VL1-CH1-CL-Fc;
VL1-VL2-VH2-VH1-CL-CH1-Fc;
VH1-VH2-VL2-VL1-CL-CH1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-L6-Fc;
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-L6-Fc;
VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-L6-Fc; or
VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-L6-Fc;
wherein the second polypeptide has a structure represented by:
VL3-VL4-VH4-VH3-Fc;
VH3-VH4-VL4-VL3-F c;
VL3-VL4-VH4-VH3-CH1-Fc;
VH3-VH4-VL4-VL3-CH1-F c;
VL3-VL4-VH4-VH3-CL-Fc;
VH3-VH4-VL4-VL3-CL-F c;
VL3-VL4-VH4-VH3-CH1-CL-Fc;
VH3-VH4-VL4-VL3-CH1-CL-F c;
VL3-VL4-VH4-VH3-CL-CH1-Fc;
VH3-VH4-VL4-VL3-CL-CH1-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-Fc;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CH1-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CH1-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CL-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CL-Fc;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CH1-L11-CL-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CH1-L11-CL-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CL-L11-CH1-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CL-L11-CH1-F c;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CH1-L11-CL-L12-Fc;
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CH1-L11-CL-L12-Fc;
VL3-L7-VL4-L8-VH4-L9-VH3-L10-CL-L11-CH1-L12-Fc; or
VH3-L7-VH4-L8-VL4-L9-VL3-L10-CL-L11-CH1-L12-F c; and
wherein:
VL1 is a first immunoglobulin light chain variable region;
VL2 is a second immunoglobulin light chain variable region;
VL3 is a third immunoglobulin light chain variable region;
VL4 is a fourth immunoglobulin light chain variable region;
VH1 is a first immunoglobulin heavy chain variable region;
VH2 is a second immunoglobulin heavy chain variable region;
VH3 is a third immunoglobulin heavy chain variable region;
VH4 is a fourth immunoglobulin heavy chain variable region;
Fc is a region comprising an immunoglobulin heavy chain constant region 2 (CH2), an immunoglobulin heavy chain constant region 3 (CH3), and optionally, an immunoglobulin hinge;
CH1 is an immunoglobulin heavy chain constant region 1;
CL is an immunoglobulin light chain constant region; and
L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11 and L12 are amino acid linkers.
131 . The antigen binding polypeptide complex of claim 128 , wherein VH1, VL1, VH3 and VL3 bind to the same antigen.
132 . The antigen binding polypeptide complex of claim 128 , wherein VH2, VL2, VH4 and VL4 bind to the same antigen.
133 . The antigen binding polypeptide complex of claim 128 , wherein VH1, VH2, VH3 and VH4 each bind to different antigens, and VL1, VL2, VL3 and VL4 each bind to different antigens.
134 . The antigen binding polypeptide complex of claim 128 , wherein linkers L1, L2, L3, L4, L5, L6, L7 and L8 each comprise the amino acid sequence of any one of SEQ ID NOs:1-19 and 681-688, or a sequence having at least 95% identity to any one of SEQ ID NOs:1-19 and 681-688.
135 . The antigen binding polypeptide complex of claim 129 , wherein linkers L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11 and L12 each comprise the amino acid sequence of any one of SEQ ID NOs:1-19 and 681-688, or a sequence having at least 95% identity to any one of SEQ ID NOs:1-19 and 681-688.
136 . The antigen binding polypeptide complex of claim 130 , wherein linkers L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11 and L12 each comprise the amino acid sequence of any one of SEQ ID NOs:1-19 and 681-688, or a sequence having at least 95% identity to any one of SEQ ID NOs:1-19 and 681-688.
137 . The antigen binding polypeptide complex of claim 128 , wherein the Fc region comprises at least one knob-into-hole modification.
138 . The antigen binding polypeptide complex claim 129 , wherein the Fc region comprises at least one knob-into-hole modification.
139 . The antigen binding polypeptide complex of claim 130 , wherein the Fc region comprises at least one knob-into-hole modification.
140 . An antibody or antigen binding fragment thereof comprising the antigen binding polypeptide complex of claim 128 .
141 . The antibody or antigen binding fragment thereof of claim 140 , wherein the antigen binding fragment is a Fab, scFab, Fab′, F(ab′)2, Fv, or scFv.
142 . The antibody or antigen binding fragment thereof of claim 140 , wherein the antibody is human or humanized.
143 . A polypeptide having at least 95% identity to any one of SEQ ID NOs:32-43, 62-79, 130-138, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, and 679, or having at least 95% identity to the amino acid sequence of SEQ ID NOs:32 or 33 that does not contain the eight histidine residues at the C-terminus.
144 . A polynucleotide having at least 95% identity to any one of SEQ ID NOs:44-55, 80-97, 139-147, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, and 680.
145 . A vector comprising the polynucleotide of claim 144 .
146 . A host cell comprising the polynucleotide of claim 144 .
147 . A pharmaceutical composition comprising (i) the antigen binding polypeptide complex of claim 128 , and (ii) a pharmaceutically acceptable carrier.
148 . A pharmaceutical composition comprising (i) a polynucleotide encoding the antigen binding polypeptide complex of claim 128 , and (ii) a pharmaceutically acceptable carrier.
149 . A method of treating or preventing a cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the antigen binding polypeptide complex of claim 128 .
150 . A method of treating or preventing a virus infection, comprising administering to a subject in need thereof a therapeutically effective amount of the antigen binding polypeptide complex of claim 128 .