Phage Display: A Practical Approach (Practical Approach Series, 266)

Phage display has become established as a powerful protein engineering method for identifying polypeptides with novel properties, and altering the properties of existing ones. Although the technique is widely used in biological research and drug discovery, it remains technically challenging, and new applications and procedures continue to evolve. Phage Display - A Practical Approach is an up-to-date, comprehensive and integrated experimental guide to the technique, useful for novice and expert alike. The book aims to enable researchers to design and undertake all aspects of a phage display project, from designing an experimental strategy and constructing a library to performing selections and analyzing the results. An introductory chapter provides an overview of phage biology and phage display, including guidelines for planning a successful phage display experiment. Individual chapters provide protocols for constructing libraries using oligonucleotide-directed mutagenesis or D.N.A. recombination, performing binding selections, and analyzing the binding activities of selected phage clones. Separate chapters then cover common applications, including selection of ligands from peptide libraries, generation of phage antibody libraries and isolation and optimization of antibodies, selection of D.N.A. binding proteins, and expression cloning using cD.N.A. display. Further chapters describe alternative selection strategies, such as selection using immune sera, selection based on enzymatic activity or protein stability, and selection in vivo. Protocols and chapters are extensively cross-referenced, allowing readers to move beyond the specific examples given to customize the procedures to their own protein or selection system of interest. Written by experts in the field, Phage Display - A Practical Approach provides a comprehensive guide to the design and execution of phage display projects, for all those using the technique i