Improved antibody-payload conjugates (apcs) prepared by site-specific conjugation utilizing genetic code expansion

A site-selectively modified immunoglobulin molecule, comprising at least one immunoglobulin heavy chain (IgH) and at least one immunoglobulin light chain (IgL),

• said IgH comprising a variable region VH encompassing
• a CDR-H1 selected from SEQ ID NO:
• 9 and 10,
• a CDR-H2 selected from SEQ ID NO:
• 11 and 12, and
• a CDR-H3, selected from SEQ ID NO:
• 13 and 14, and a constant region CH;
• and
• said IgL comprising a variable region VL encompassing
• a CDR-L1 selected from SEQ ID NO:
• 15 and 16,
• a CDR-L2 selected from SEQ ID NO:
• 17 and 18 and
• a CDR-L3 selected from SEQ ID NO:
• 19 and 20;
• and
• a constant region CL;
• comprising
• at least one side-selectively modified IgH;
• or
• at least one side-selectively modified IgL;
• or
• at least one side-selectively modified IgH and at least one side-selectively modified IgL;
• wherein
• said side-selectively modified immunoglobulin molecule has the ability to bind human epidermal growth factor receptor 2 (ERBB2 or HER2/neu);
• said site-selectively modified immunoglobulin molecule is present in non-glycosylated, glycosylated or de-glycosylated form;
• and
• said side-selectively modified immunoglobulin molecule comprises one site-selective modification in IgH and/or IgL, wherein said one site-selective modification is selected from
• a site-selectively modified IgH comprising an ncAA in one amino acid
• sequence position corresponding to a position selected from
• a) VH positions:
• S25, P41, G42, K43, R50, D62, K65, E89, D102;
• b) CH1 positions:
• E155, P156, S194, E219;
• c) CH2 positions:
• K249, K251, E275, K277, D283, H288, K291, K293, E296, R304, K320, K323 and K343
• each of SEQ ID NO:
• 2,
• and/or
• a site-selectively modified IgL comprising an ncAA in one amino acid sequence position corresponding to a position selected from
• d) VL positions:
• G41, K42, K45, A51, P59, R61, D70, E81;
• e) CL positions:
• A111, E143, D151, G157, K169, G200
• each of SEQ ID NO:
• 4;
• or
• said side-selectively modified immunoglobulin molecule comprises two site-selective modifications, wherein said two site-selective modifications are selected from
• double modifications in two amino acid sequence positions of at least one IgH corresponding to positions selected from:
• a) CH1 positions:
• E155, P156, S194, E219;
• b) CH2 positions:
• K249, K251, E275, K277, D283, H288, K291, K293, E296, R304, K320, K323 and K343
• each of SEQ ID NO:2;
• or
• double modifications of two amino acid sequence positions in at least one IgL corresponding to positions selected from:
• c) CL positions:
• A111, E143, D151, G157, K169, G200 each of SEQ ID NO:4.