Antibody Humanization

The goal of antibody humanization is to engineer a monoclonal antibody (mAb) raised in a nonhuman species into one that is less immunogenic when administered to humans.

The complementarity-determining regions (CDR) grafting method in which the antigen-binding specificity of a non-human antibody is transferred to a human antibody by replacing the CDR loops and selected variable-region framework residues (FRs).

CDR grafting has three fundamental components:

  1. Delimitation of CDRs as regions determining the specificity of the nonhuman antibody and hence target for grafting
  2. Selection of human FRs to be combined with the nonhuman CDRs
  3. Assessment of conflicts between nonhuman CDRs and human FRs and design of back mutations to prevent a loss of affinity in the final products.

Material Requirements:

1      Antibody with desired specificity from any parental species (eg. mouse, rat, rabbit, camel).

2      Hybridoma cells (mouse, rabbit, et al.) or sequences of antibody: VL and VH.

3      Antigen and assay protocol for measuring binding specificity (ELISA, Western blot) and affinity of humanized antibody by Octet.

antibody humanization modellingHomology modelling of mouse and humanization template sequence using Rosetta homology modelling server.