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

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.

  "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

  "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

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