Generation of Naive Human Antibody Libraries

Abstract

The phage display of antibody fragments as Fabs or scFvs (McCafferty et al., 1990), has its origins in experiments demonstrating that both small peptides and folded proteins could be displayed on the surface of filamentous bacteriophage (Smith, 1985; Bass et al., 1990). Since the generation of the first human antibodies by phage display (Winter et al., 1994), the technology has developed to the point where large scFv repertoires have been created that yield antibodies with sub-nanomolar affinities (Vaughan et al., 1996; Xie et al., 1997). This is comparable with the best antibodies obtained using hybridoma technology. Phage display repertoires can also be used to isolate antibodies not easily obtained by hybridoma technology, such as those specific for toxic proteins and human anti-self antibodies (Griffiths et al., 1993; Vaughan et al., 1998). In addition, using a variety of selection and screening strategies, the same non-immunised “single pot“ library can be used to simultaneously derive many high affinity antibodies with different specificities in only a matter of weeks. Furthermore, selection techniques have evolved to the point where strategies now exist that facilitate selections on complex antigens expressed on the cell surface (Chapter 12).