14 - Construction, Screening and Expression of Recombinant Antibodies

Abstract

This chapter focuses on the use of bacteria and bacteriophages to display a diverse library of antibodies equivalent to the mammalian immune repertoire. The new technology combines production, selection, and affinity maturation in numerous imaginative ways. It is, however, rather more complex than cell fusion, and still has some technical and logistical problems. None the less, it holds great promise, especially for the production of human monoclonal antibodies. The chapter presents a representative overview of the phagedisplay technology and some of the processes by which phage-libraries are made and handled. There are many attractions to the production of antibodies in E. coli. The problems of production of human antibodies are largely overcome by phage display and expression in bacteria, although many problems remain. Manipulation of sequences for altering the affinity and specificity is possible by random or site-directed mutagenesis. It is relatively straightforward to add extra sequences as tags or for effector functions.