Phage Display A Practical Approach

by Clackson, Tim; Lowman, Henry B.

• ISBN13: 9780199638741
• ISBN10: 0199638748
• Edition: 1st
• Format: Hardcover
• Copyright: 2004-05-06
• Publisher: Oxford University Press

Summary

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 DNA 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 DNA binding proteins, and expression cloning using cDNA 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 in basic research and drug discovery.

Table of Contents

Introduction to phage biology and phage display, Russel, Lowman and Clackson

Constructing phage display libraries by oligonucleotide-directed mutagenesis, Sidhu and Weiss

In vitro DNA recombination, Miyazaki and Arnold

Phage selection strategies for improved affinity and specificity of proteins and peptides, Dennis and Lowman

Rapid screening of phage displayed protein binding affinities by phage ELISA, DeLano and Cunningham

Identification of novel ligands for receptors using recombinant peptide libraries, Cwirla et al

Substrate phage display, Matthews and Ballinger

Protease-based selection of stably folded proteins and protein domains from phage display libraries, Tuna et al

Phage display of zinc fingers and other nucleic acid-binding motifs, Isalan and Choo

In vivo and ex vivo selections using phage-displayed libraries, Hoffman et al

Screening phage libraries with sera, Monaci and Cortese

Interaction cloning using cDNA libraries displayed on phage, Jespers and Fransen

Phage antibody libraries, Bradbury and Marks

Affinity maturation of phage antibodies, Neilson and Marks

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