CPC C07K 16/2809 (2013.01) [C07K 16/2848 (2013.01); C07K 16/2863 (2013.01); C07K 16/2866 (2013.01); C07K 16/2875 (2013.01); C07K 16/2896 (2013.01); C07K 16/30 (2013.01); C07K 16/4283 (2013.01); C07K 2317/24 (2013.01); C07K 2317/31 (2013.01); C07K 2317/526 (2013.01); C07K 2317/55 (2013.01); C07K 2317/565 (2013.01); C07K 2317/76 (2013.01); C07K 2319/00 (2013.01); C07K 2319/70 (2013.01)] | 18 Claims |
1. A method of selecting an antigen-binding molecule that recognizes a first antigen that is human CD3 and a second antigen different from the first antigen, the method comprising:
identifying a starting antigen-binding molecule comprising a starting antibody variable region comprising a VH and a VL that together recognize the first antigen and do not recognize the second antigen;
preparing a library of altered antigen-binding molecules comprising variable regions that vary from the starting antibody variable region and from each other by at least one amino acid substitution or insertion at a position or positions located within any one or more of a VH CDR1, a VH CDR2, a VH CDR3, a VH FR3 region, a VL CDR1, a VL CDR2, or a VL CDR3; and
selecting from the library an altered antigen-binding molecule comprising an altered antibody variable region comprising a first antigen-binding site and a second antigen-binding site, wherein:
(a) the first antigen-binding site recognizes the first antigen; and
(b) the second antigen-binding site recognizes the second antigen, wherein the second antigen is selected from an Fcγ receptor (FcγR), a toll-like receptor (TLR), a lectin, an IgA, an immune checkpoint molecule, a tumor necrosis factor (TNF) superfamily molecule, a TNF receptor (TNFR) superfamily molecule, and an NK receptor molecule; and
(c) the first antigen-binding site cannot bind to the first antigen when the second antigen-binding site is bound to the second antigen; and
(d) the second antigen-binding site cannot bind to the second antigen when the first antigen-binding site is bound to the first antigen,
thereby selecting an antigen-binding molecule that recognizes the first antigen and the second antigen.
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