Generation and Panning of a Naïve Human scFv Phage Display Library

Antibody libraries are the heartbeat of any phage display campaign to isolate monoclonal antibodies. Naïve antibody libraries have been used to identify monoclonal antibodies against a collection of different targets from small molecules to large protein targets. The fundamental requirements of a good naïve library generation is the availability of a diverse collection of antibody genes as the genetic source. The antibody gene pool used for naïve antibody libraries is the IgM repertoire derived from healthy donors. The obtained antibody genes will then be cloned as single-chain fragment variable (scFv), as it is a common format used for its smaller size and preference for phage display. The protocol will encompass the library cloning process which involves the use of a two-step cloning method, followed by bio-panning selection.