Selection by Phage Display of Single Domain Antibodies Specific to Antigens in Their Native Conformation

Abstract

Phage display of antibody fragments and other binding molecules is a well-established technique to identify ligands interacting with any molecule of interest. Selection of in vivo matured single domain antibody fragments from phage display libraries is very powerful as in these libraries each clone represents a noncombinatorial functional domain of a naturally circulating antibody, and thus such libraries contain a high number of antigen-specific clones. Consequently, individual binders to antigens of interest are efficiently obtained typically after one or two selection rounds. Furthermore, the large functional diversity within these antibody libraries allows the application of different and more stringent selection conditions resulting in the selection of complementary antibody panels. In this chapter, we present a guide to perform selections against purified antigens and antigens in their native conformation and context.