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Phage Display: A Laboratory Manual


Subject Area(s):  Molecular BiologyLaboratory Techniques

By Carlos F. Barbas III, The Scripps Research Institute; Dennis R. Burton, The Scripps Research Institute; Jamie K. Scott, Simon Fraser University; Gregg J. Silverman, University of California, San Diego

© 2001 • 736 pp., illus., index
This book has been produced using print on demand technology.
Paperback • $81.00 32.40
ISBN  978-087969740-2
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  •     Description    
  •     Contents    
  •     Reviews    

Description

Phage-display technology has begun to make critical contributions to the study of molecular recognition. DNA sequences are cloned into phage, which then present on their surface the proteins encoded by the DNA. Individual phage are rescued through interaction of the displayed protein with a ligand, and the specific phage is amplified by infection of bacteria.

Phage-display technology is powerful but challenging and the aim of this manual is to provide comprehensive instruction in its theoretical and applied so that any scientist with even modest molecular biology experience can effectively employ it. The manual reflects nearly a decade of experience with students of greatly varying technical expertise andexperience who attended a course on the technology at Cold Spring Harbor Laboratory.

Phage-display technology is growing in importance and power. This manual is an unrivalled source of expertise in its execution and application.

Contents

SECTION 1 – PHAGE DISPLAY
Chapter 1.   Filamentous Phage Biology
Chapter 2.   Phage-display Vectors
Chapter 3.   Antibody Libraries
Chapter 4.   Peptide Libraries
Chapter 5.   Functional Domains and Scaffolds
Chapter 6.   Gene Fragment Libraries and Genomic and cDNA Expression Cloning
SECTION 2 – ANTIBODY LIBRARIES
Chapter 7.   Overview: Amplification of Antibody Genes
Chapter 8.   Generation of Antibody Libraries: Immunization, RNA Preparation, and cDNA Synthesis
Chapter 9.   Generation of Antibody Libraries: PCR Amplification and Assembly of Light- And Heavy-chain Coding Sequences
Chapter 10.   Selection from Antibody Libraries
Chapter 11.   Analysis of Selected Antibodies
Chapter 12.   Production and Purification of Fab and scFv
Chapter 13.   Antibody Engineering
SECTION 3 – PEPTIDE LIBRARIES
Chapter 14.   Overview: Peptide Libraries
Chapter 15.   General Phage Methods
Chapter 16.   Production of Peptide Libraries
Chapter 17.   Screening Peptide Libraries
Chapter 18.   Analysis of Phage-borne Peptides
Chapter 19.   Construction and Use of pIII-displayed Peptide Libraries
SECTION 4 – OTHER LIBRARIES AND OTHER METHODS OF PANNING
Chapter 20.   Construction and Selection from Gene Fragment Phage-display Expression Libraries
Chapter 21.   Construction and Selection from cDNA Phage-display Expression Libraries
Chapter 22.   In Vivo Selection of Phage-display Libraries
Chapter 23.   Cell-surface Selection and Analysis of Monoclonal Antibodies from Phage Libraries
SECTION 5 – APPENDICES
Appendix 1.   Useful Information
Appendix 2.   Recipes
Appendix 3.   General Procedures
Appendix 4.   Cautions
Appendix 5.   Suppliers
Appendix 6.   Trademarks

Reviews

review:  "The book's raison d'être is as a source of detailed protocols. These are clearly written, and include ideas for experimental controls and useful trouble-shooting notes. Jargon is generally avoided, and the protocols are up to date. …[This] is a much-needed and well-executed attempt at theoretical and practical support to any researcher, established or novice, using phage display."
      —Trends in Genetics

review:  "This book is a solid work that addresses both theoretical and practical aspects of phage display technology. It covers the use of phage display in protein engineering with particular emphasis on antibody, peptide, cDNA libraries and ligand receptor engineering, which together represent [approximately] 90% of the phage display field. Anyone using or contemplating any of these applications would find it worthwhile to read the relevant chapters."
      —Trends in Biotechnology