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. 2024 Jul 16;13(3):57. doi: 10.3390/antib13030057

Figure 1.

Figure 1

Humanization by CDR grafting is insufficient to retain affinity to HLA-DR. (A) Parental 44H10 heavy and light chain amino acid sequences annotated with IMGT (blue) and Kabat (orange) CDR definitions. The IMGT unique numbering of key conserved amino acids is provided in blue above each sequence. (B) Conventional CDR-grafting process used to generate humanized 44H10 versions 1–18, grafting IMGT- or Kabat-defined CDRs onto selected human antibody frameworks. (C,D) Flow cytometry screening of humanized 44H10 V1–9 (C) and V10–18 (D) binding to BJAB cells (10 µg/mL concentration). The candidate showing modest antigen binding (V17) is colored in green. (E) Flow cytometry titration curves of parental 44H10 and V17 IgGs binding to BJAB cells. (F) BLI binding profile of parental 44H10 and V17 IgGs to recombinant HLA-DR, where black lines represent measured binding and gray/green curves correspond to the data fitted to a 1:1 binding model.