Phage Display: A Practical Approach

ISBN-13: 9780199638734 / Angielski / Miękka / 2004 / 360 str.

Tim Clackson; Henry B. Lowman

Opis

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

O autorze

Dr Tim Clackson Vice President, Gene Therapy and Genomics ARIAD Pharmaceuticals, Inc., 26 Landsdowne Street, Cambridge, MA 02139, USA +1 617 494 0400 ext. 258 +1 617 225 2589 clackson@ariad.com Tim Clackson, Ph.D. is Senior Vice President, Science and Technology at ARIAD Pharmaceuticals, Inc., where he has served for eight years in a variety of research and research management positions. His group is combining chemistry, cell biology and protein engineering to develop techniques for regulating gene expression and other intracellular processes using small molecules. Dr Clackson's particular research interest is the use of protein engineering and chemical biology techniques to analyze and redesign the interfaces between molecules, especially small molecule-protein interfaces. Dr Henry B. Lowman Senior Scientist Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA +1 650 225 1171 +1 650 225 3734 hbl@gene.com Henry B. Lowman, Ph.D. is a Senior Scientist and Associate Director of Antibody Engineering with joint appointments in Immunology and Protein Engineering, at Genentech, Inc., where he has been engaged in research on proteins and peptides for over thirteen years.