CLONING IN A BACTERIOPHAGE-LAMBDA VECTOR FOR THE DISPLAY OF BINDING-PROTEINS ON FILAMENTOUS PHAGE

We have combined the efficiency and ease of use of bacteriophage h vec tors with the power of phage display screening technology to create Su rfZAP(TM). The use of bacteriophage h allows the construction of large h expression libraries, which are rapidly and efficiently converted t o stable plasmid libraries by mass excision. In SurfZAP, clones are ex pressed as fusions with amino acids 198-406 of the M13 minor coat prot ein (cpIII) and are displayed on the surface of filamentous phage. Whe n produced with helper phage proteins, the fusion proteins are incorpo rated into the surface of phagemid particles. We demonstrate the utili ty of biopanning by isolating tetanus toxoid-binding mouse Fab clones from SurfZAP libraries. Approximately 10-100-fold enrichment of specif ic clones was observed after each panning round. The ability to create a large library of genotypes and screen the phenotypes by activity ma y be a potent methodology for basic research and drug discovery.