Antibody humanization aims to reduce the immunogenicity of monoclonal antibodies from xenogeneic origin by replacing non-human framework regions, (FR), with human ones. It is a crucial step in therapeutic antibody development.
- Sequencing of the VH and VL genes
- Modelling of the parent VH and VL domains
- Alignment with a range of preferred human germline sequences
- Assessment of conflicts between non-human CDRs and human FRs and design of back mutations to prevent a loss of affinity in the final products.
- CDR grafting onto preferred germline backbones
- ~8 different humanized sequences generated
- Synthesis, cloning and small-scale production of all humanized variants, original parent and chimeric controls in our mammalian expression system
- Characterization of the variants by ELISA or SPR
- Detailed report
- ~14 weeks from reception of the VH and VL sequences
Robert R, Streltsov VA, Newman J, Pearce LA, Wark KL, Dolezal O. Germline humanization of a murine Abeta antibody and crystal structure of the humanized recombinant Fab fragment. Protein Sci. 2010: 19:299-308.