Design and generation of synthetic antibody libraries for phage display

Highly functional synthetic antibody libraries can be used to generate antibodies against a multitude of antigens with affinities and specificities that rival or exceed those of natural antibodies. Current design and generation of synthetic antibody libraries are dependent on our insights from previous studies of simplified synthetic antibody libraries, in addition to our knowledge of antibody structure and function and sequence diversity of natural antibody repertoires. We describe a detailed protocol for the design and generation of phage-displayed synthetic antibody libraries built on a single framework with diversity restricted to four complementarity-determining regions by using precisely designed degenerate oligonucleotides. This general methodology could be applied to generation of large, functional synthetic antibody libraries using standard supplies, equipment, and molecular biology techniques.