Generation and Selection of Rabbit Antibody Libraries by Phage Display
The rabbit antibody repertoire is an exceptional source for both polyclonal antibodies (pAbs) and monoclonal antibodies (mAbs) that combine high specificity with high avidity and affinity, respectively. In contrast to rabbit pAbs, which have been utilized for decades, rabbit mAbs are defined reagents with infinite supply, prompting considerable interest in recent years. Phage-display technology based on chimeric rabbit/human Fab libraries allows the selection of rabbit mAbs that cross-react with mouse and human antigens while maintaining high affinity and specificity.
Key wordsRabbit monoclonal antibodies phage display phagemid antibody library Fab panning
I would like to thank Dr. Carlos F. Barbas III (The Scripps Research Institute, La Jolla, CA) for training me in antibody technologies including phage display, and members of my laboratory at the National Cancer Institute, in particular Drs. Thomas Hofer and Ka Yin Kwong, for their outstanding contributions to further evolving the methodology.
- 5.Rader, C., Steinberger, P., and Barbas, C. F., III (2001) Selection from antibody libraries, in Phage Display: A Laboratory Manual (Barbas, C. F., III, Burton, D.R., Scott, J. K., and Silverman, G. J., eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
- 6.Rader, C., Ritter, G., Nathan, S., Elia, M., Gout, I., Jungbluth, A. A., Cohen, L. S., Welt, S., Old, L. J., and Barbas, C. F., III (2000) The rabbit antibody repertoire as a novel source for the generation of therapeutic human antibodies. J. Biol. Chem. 275, 13668–13676.PubMedCrossRefGoogle Scholar
- 7.Popkov, M., Mage, R. G., Alexander, C. B., Thundivalappil, S., Barbas, C. F., III, and Rader, C. (2003) Rabbit immune repertoires as sources for therapeutic monoclonal antibodies: the impact of kappa allotype-correlated variation in cysteine content on antibody libraries selected by phage display. J. Mol. Biol. 325, 325–335.PubMedCrossRefGoogle Scholar
- 9.Hofer, T., Tangkeangsirisin, W., Kennedy, M. G., Mage, R. G., Raiker, S. J., Venkatesh, K., Lee, H., Giger, R. J., and Rader, C. (2007) Chimeric rabbit/human Fab and IgG specific for members of the Nogo-66 receptor family selected for species cross-reactivity with an improved phage display vector. J. Immunol. Methods 318, 75–87.PubMedCrossRefGoogle Scholar
- 11.Andris-Widhopf, J., Steinberger, P., Fuller, R., Rader, C., and Barbas, C. F., III (2001) Generation of antibody libraries: PCR amplification and assembly of light- and heavy-chain coding sequences, in Phage Display: A Laboratory Manual (Barbas, C. F., III, Burton, D.R., Scott, J. K., and Silverman, G. J., eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
- 13.Scott, J. K. and Barbas, C. F., III (2001) Phage-display vectors, in Phage Display: A Laboratory Manual (Barbas, C. F., III, Burton, D.R., Scott, J. K., and Silverman, G. J., eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
- 14.Barbas, C. F., III, Kang, A. S., Lerner, R. A., and Benkovic, S. J. (1991) Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc. Natl. Acad. Sci. USA 88, 7978–7982.Google Scholar