| CPC C07K 16/32 (2013.01) [A61P 35/00 (2018.01); G01N 33/57415 (2013.01); G01N 33/57446 (2013.01); G01N 33/57492 (2013.01); C07K 2317/24 (2013.01); C07K 2317/31 (2013.01); C07K 2317/35 (2013.01); C07K 2317/526 (2013.01); C07K 2317/55 (2013.01); C07K 2317/565 (2013.01); C07K 2317/567 (2013.01); C07K 2317/64 (2013.01); C07K 2317/73 (2013.01); C07K 2317/77 (2013.01); C07K 2317/92 (2013.01); C07K 2317/94 (2013.01); G01N 2333/71 (2013.01)] | 19 Claims |
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1. An antigen-binding construct comprising a variant first antigen-binding polypeptide construct which monovalently binds a first HER2 ECD2 (human epidermal growth factor receptor 2 extracellular domain 2) antigen, the variant first antigen-binding polypeptide construct comprising
a variable heavy (VH) domain comprising a complementary determining region (CDR) 1 (CDR-H1) comprising the sequence as set forth in SEQ ID NO: 956, a CDR-H2 comprising the sequence as set forth in SEQ ID NO: 957, and a CDR-H3 comprising the sequence as set forth in SEQ ID NO: 958; and
a variable light (VL) domain comprising a CDR-L1 comprising the sequence as set forth in SEQ ID NO: 959, a CDR-L2 comprising the sequence as set forth in SEQ ID NO: 960, and a CDR-L3 comprising the sequence as set forth in SEQ ID NO: 609;
and wherein the Glycine at position 56 of CDR-H2 has been substituted with a Tyrosine (H_G56Y) or a Phenylalanine (H_G56F), or the Serine at position 99 of CDR-H3 has been substituted with a Tryptophan (H_S99W), numbering according to Kabat numbering system;
optionally wherein the variant first antigen-binding polypeptide construct comprises H_G56Y and further comprises the following substitution or set of substitutions relative to the VH domain as set forth in SEQ ID NO: 2 and the VL domain as set forth in SEQ ID NO: 11, numbering according to Kabat numbering system;
H_K75W; or
H_T30Q; or
H_T30Y; or
H_S99W; or
L_Y49W; or
L_Y96G; or
H_S99W and L_Y49W; or
L_Y49W and L_Y96G; or
H_T30Q and L_Y49W; or
H_T30Q and H_S99W; or
H_T30Q and L_Y96G; or
H_T30Y and L_Y49W; or
H_T30Q and H_S99W and L_Y49W; or
H_T30Q and L_Y49W and L_Y96G; and
optionally wherein the variant first antigen-binding polypeptide construct comprises H_S99W and further comprises the following substitution or set of substitutions relative to the VH domain as set forth in SEQ ID NO: 2 and the VL domain as set forth in SEQ ID NO: 11, numbering according to Kabat numbering system;
H_K75W; or
H_T30Q; or
H_K75E; or
H_T30Y; or
H_K75W and L_Y49W; or
H_T30Q and H_K75W; or
H_T30Q and L_Y49W; or
H_T30Q and H_K75W and L_Y49W; or
H_K75W and L Y49W and L_Y96G; or
H_T30Q and H_K75W and L_Y96G; or
H_T30Q and L_Y49W and L_Y96G; or
H_T30Q and H_G56Y and L_Y49W.
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