The rabbit antibody repertoire as a novel source for the generation of therapeutic human antibodies

The rabbit antibody repertoire, which in the form of polyclonal antibodies has been used in diagnostic applications for decades, would be an attractiv e source for the generation of therapeutic human antibodies. The humanizati on of rabbit antibodies, however, has not been reported. Here we use phage display technology to select and humanize antibodies from rabbits that were immunized with human A33 antigen which is a target antigen for the immunot herapy of colon cancer. We first selected rabbit antibodies that bind to a cell surface epitope of human A33 antigen with an affinity in the 1 nM rang e. For rabbit antibody humanization, we then used a selection strategy that combines grafting of the complementarity determining regions with framewor k fine tuning. The resulting humanized antibodies were found to retain both high specificity and affinity for human A33 antigen.