Humanization of chicken monoclonal antibody using phage-display system.
We describe a simple method for humanizing chicken monoclonal antibody (mAb). Humanization of mAbs by simple CDR-grafting often results in loss of affinity because certain framework residues of the antibody variable regions can participate in antigen-antibody interaction. In this study, humanization of chicken mAbs was achieved by CDR-grafting, followed by framework fine-tuning using a chicken phage-displayed mAb, phAb4-31, as a model antibody. In order to fine-tune the framework, we used the phage-displayed combinatorial library with permutation of important framework residues. After panning the humanized library, the "most humanized" variants were selected and analyzed for antigen-binding activity. All of these clones retained affinity comparable to the parental chicken mAb. These results suggest that chicken mAbs can easily be humanized, and thus humanized chicken mAbs may be practically applied as therapeutic agents.
Nishibori
Nahoko
N
Laboratory of Immunobiology, Department of Molecular and Applied Bioscience, Graduate School of Biosphere Science, Hiroshima University, 1-4-4 Kagamiyama, Higashi-Hiroshima 739-8528, Japan.
Horiuchi
Hiroyuki
H
Furusawa
Shuichi
S
Matsuda
Haruo
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics*; D000915:Antibody Affinity; D002645:Chickens; D003001:Cloning, Molecular; D022801:Complementarity Determining Regions / Q000235:genetics; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D008722:Methods; D019151:Peptide Library; D015202:Protein Engineering
D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library