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 attractive source for the generation of therapeutic human antibodies. The humanization 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 immunotherapy 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 range. For rabbit antibody humanization, we then used a selection strategy that combines grafting of the complementarity determining regions with framework fine tuning. The resulting humanized antibodies were found to retain both high specificity and affinity for human A33 antigen.
Rader
C
C
Skaggs Institute for Chemical Biology, Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Ritter
G
G
Nathan
S
S
Elia
M
M
Gout
I
I
Jungbluth
A A
AA
Cohen
L S
LS
Welt
S
S
Old
L J
LJ
Barbas
C F
CF
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology / Q000627:therapeutic use*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D015723:Gene Library; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D011817:Rabbits; D013045:Species Specificity
D000906:Antibodies; C104258:GPA33 protein, human; D008562:Membrane Glycoproteins