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Advances in Phage Display, A Laboratory Manual


Subject Area(s):  Biotechnology; Molecular Biology; Laboratory Techniques

Edited by Gregg J. Silverman, New York University School of Medicine; Christoph Rader, The Scripps Research Institute; Sachdev S. Sidhu, University of Toronto

Download a Free Excerpt from Advances in Phage Display: A Laboratory Manual:

Preface
Advances in Phage Display—A Perspective
Index

Due March 2026 • 472 pages, illustrated (63 color and 17 B&W), index
Paperback •
ISBN  978-1-621824-25-1

  •     Description    
  •     Contents    

Description

Phage display is a powerful technique that has greatly accelerated progress in the fields of drug discovery, structural biology, biomarker development, identification of antigenic targets, and protein domain engineering. This technology leverages bacteriophages—viruses that infect bacteria—to display a wide range of foreign peptides, proteins, or antibody fragments on their surfaces, allowing for the identification of optimal binding molecules.

With contributions from instructors and former students from over three decades of the “Antibody Engineering and Display Technologies” Cold Spring Harbor Laboratory course, this collection provides expert reviews on principles and applications of phage display, along with detailed laboratory protocols, and represents an essential resource for those using phage display in molecular biology and biotechnology.

Contents

Preface
SECTION 1. FOUNDATIONS OF PHAGE DISPLAY
CHAPTER 1 OVERVIEW
Advances in Phage Display—A Perspective
George P. Smith
CHAPTER 2 OVERVIEW
Structure, Biology, and Applications of Filamentous Bacteriophages
Jasna Rakonjac, Vicki A.M. Gold, Rayén I. León-Quezada, and Catherine H. Davenport
CHAPTER 3 OVERVIEW
Principles of Affinity Selection
George P. Smith
CHAPTER 4 OVERVIEW
The pComb3 Phagemid Family of Phage Display Vectors
Christoph Rader
SECTION 2. POSTIMMUNE ANTIBODY PHAGE DISPLAY LIBRARIES
CHAPTER 5 INTRODUCTION
Generation and Selection of Phage Display Antibody Libraries in Fab Format
Christoph Rader
PROTOCOLS
1 Generation of Antibody Libraries for Phage Display: Human Fab Format
Haiyong Peng and Christoph Rader
2 Generation of Antibody Libraries for Phage Display: Chimeric Rabbit/Human Fab Format
Haiyong Peng and Christoph Rader
3 Generation of Antibody Libraries for Phage Display: Preparation of Electrocompetent E. coli
Haiyong Peng and Christoph Rader
4 Generation of Antibody Libraries for Phage Display: Preparation of Helper Phage
Haiyong Peng and Christoph Rader
5 Generation of Antibody Libraries for Phage Display: Library Reamplification
Haiyong Peng and Christoph Rader
6 Phage Display Selection of Antibody Libraries: Panning Procedures
Haiyong Peng and Christoph Rader
7 Phage Display Selection of Antibody Libraries: Screening of Selected Binders
Haiyong Peng and Christoph Rader
8 Cloning, Expression, and Purification of Phage Display-Selected Fab for Biophysical and Biological Studies
Matthew G. Cyr, Haiyong Peng, and Christoph Rader
SECTION 3. CHICKEN ANTIBODY PHAGE DISPLAY LIBRARIES
CHAPTER 6 INTRODUCTION
Generation of Chicken Antibody Libraries and Selection of Antigen Binders
Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, and Junho Chung
PROTOCOLS
1 Chicken Immunization Followed by RNA Extraction and cDNA Synthesis for Antibody Library Preparation
Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, and Junho Chung
2 Generation of a Phage Display Chicken Single-Chain Variable Fragment Library
Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, and Junho Chung
3 Preparation of VCSM13 Helper Phage for Display Library Reamplification and Bio-Panning
Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, and Junho Chung
4 Selection of Antigen Binders from a Chicken Single-Chain Variable Fragment Library
Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, and Junho Chung
SECTION 4. SYNTHETIC HUMAN ANTIBODY PHAGE DISPLAY LIBRARIES
CHAPTER 7 OVERVIEW
Structural Survey of Antigen Recognition by Synthetic Human Antibodies
Maryna Gorelik, Shane Miersch, and Sachdev S. Sidhu
CHAPTER 8 INTRODUCTION
Beyond Natural Immune Repertoires: Synthetic Antibodies
Gianluca Veggiani and Sachdev S. Sidhu
PROTOCOL
1 Generation and Selection of Synthetic Human Antibody Libraries via Phage Display
Gianluca Veggiani and Sachdev S. Sidhu
SECTION 5. SEMISYNTHETIC scFv PHAGE DISPLAY LIBRARIES
CHAPTER 9 INTRODUCTION
Considerations for Using Phage Display Technology in Therapeutic Antibody Drug Discovery
Mary Ann Pohl and Juan C. Almagro
PROTOCOLS
1 Semisynthetic Phage Display Library Construction: Design and Synthesis of Diversified Single-Chain Variable Fragments and Generation of Primary Libraries
Juan C. Almagro and Mary Ann Pohl
2 Semisynthetic Phage Display Library Construction: Generation of Filtered Libraries
Juan C. Almagro and Mary Ann Pohl
3 Semisynthetic Phage Display Library Construction: Generation of Single-Chain Variable Fragment Secondary Libraries
Juan C. Almagro and Mary Ann Pohl
SECTION 6. HIGH-THROUGHPUT ANTIBODY REPERTOIRE LIBRARIES
CHAPTER 10 INTRODUCTION
Beyond Single Clones: High-Throughput Sequencing in Antibody Discovery
Ahmed S. Fahad, Matías F. Gutiérrez-Gonzalez, Bharat Madan, and Brandon J. DeKosky
PROTOCOLS
1 Clonal Variant Analysis of Antibody Engineering Libraries
Ahmed S. Fahad, Matías F. Gutiérrez-Gonzalez, Bharat Madan, and Brandon J. DeKosky
2 Antibody Data Analysis from Diverse Immune Libraries
Ahmed S. Fahad, Matías F. Gutiérrez-Gonzalez, Bharat Madan, and Brandon J. DeKosky
3 Clonal Lineage and Gene Diversity Analysis of Paired Antibody Heavy and Light Chains
Ahmed S. Fahad, Matías F. Gutiérrez-Gonzalez, Bharat Madan, and Brandon J. DeKosky
SECTION 7. B-CELL EPITOPE PHAGE DISPLAY LIBRARIES
CHAPTER 11 INTRODUCTION
Insights from the Study of B-Cell Epitopes of a Microbial Pathogen by Phage Display
Gregg J. Silverman
PROTOCOL
1 Cloning and Selection from Antigen Fragment Libraries for Epitope Identification
Gregg J. Silverman
SECTION 8. DOMAIN ENGINEERING IN PHAGE DISPLAY LIBRARIES
CHAPTER 12 OVERVIEW
Use of Phage Display and Other Molecular Display Methods for the Development of Monobodies
Akiko Koide and Shohei Koide
CHAPTER 13 OVERVIEW
Phage-Displayed SH2 Domain Libraries: From Ultrasensitive Tyrosine Phosphoproteome Probes to Translational Research
Gregory D. Martyn and Gianluca Veggiani
CHAPTER 14 OVERVIEW
Generation and Characterization of Engineered Ubiquitin Variants to Modulate the Ubiquitin Signaling Cascade
Chen T. Liang, Olivia Roscow, and Wei Zhang
CHAPTER 15 INTRODUCTION
Engineering of Affibody Molecules
Stefan Ståhl, Hanna Lindberg, Linnea Charlotta Hjelm, John Löfblom, and Charles Dahlsson Leitao
PROTOCOLS
1 Cloning of Affibody Libraries for Display Methods
Stefan Ståhl, Linnea Charlotta Hjelm, Charles Dahlsson Leitao, John Löfblom, and Hanna Lindberg
2 Selection of Affibody Molecules Using Phage Display
Linnea Charlotta Hjelm, Charles Dahlsson Leitao, Stefan Ståhl, John Löfblom, and Hanna Lindberg
3 Selection of Affibody Molecules Using Escherichia coli Display
Charles Dahlsson Leitao, Linnea Charlotta Hjelm, Stefan Ståhl, John Löfblom, and Hanna Lindberg
4 Selection of Affibody Molecules Using Staphylococcal Display
John Löfblom, Linnea Charlotta Hjelm, Charles Dahlsson Leitao, Stefan Ståhl, and Hanna Lindberg
Appendix: General Safety and Hazardous Material Information
Index