Antibody Phage Display

by Aitken, Robert

• ISBN13: 9781603273015
• ISBN10: 1603273018
• Edition: 2nd
• Format: Hardcover
• Copyright: 2009-07-07
• Publisher: Humana Pr Inc

Summary

Since its introduction almost 20 years ago, phage display technology has revolutionized approaches to the analysis of biomedical problems, quickly impacting the fields of immunology, cell biology, biotechnology, pharmacology, and drug discovery. In Antibody Phage Display: Methods and Protocols, Second Edition, expert researchers explore the latest in this cutting-edge technology, providing an invaluable resource that will guide readers in the design and execution of experiments based around antibody phage display. Chapters present a wide range of methods of isolating recombinant antibodies from phage display libraries, examine how the targets recognized by antibodies of interest can be identified, discuss the identification and exploitation of antibodies that can enter cells and bind to cytosolic targets, and include novel approaches to the expression of recombinant antibodies. Composed in the highly successful Methods in Molecular Biology'„¢ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.Detailed and innovative, Antibody Phage Display: Methods and Protocols, Second Edition is a critical handbook on phage display technology which is certain to stimulate the reader's imagination as much as it will guide future practice in the laboratory.

Table of Contents

Preface	p. v
Contributors	p. ix
Antibody Phage Display: Overview of a Powerful Technology that Has Quickly Translated to the Clinic	p. 1
Design and Construction of Synthetic Phage-Displayed Fab Libraries	p. 17
Affinity Isolation of Antigen-Specific Circulating B Cells for Generation of Phage Display-Derived Human Monoclonal Antibodies	p. 37
Construction of Phage Antibody Repertoires from the Blood of West Nile Virus-Infected Donors	p. 45
Anti-ß2 GP-I and Anti-prothrombin Antibodies Generated by Phage Display	p. 61
The Generation and Selection of Single-Domain, V Region Libraries from Nurse Sharks	p. 71
The Isolation of scFvs Against Small Target Molecules	p. 83
Display and Selection of scFv Antibodies on HEK-293T Cells	p. 99
Isolation of scFvs that Inhibit the NS3 Protease of Hepatitis C Virus by a Combination of Phage Display and a Bacterial Genetic Screen	p. 115
Guided Selection Methods Through Chain Shuffling	p. 133
Sequential Antigen Panning for Selection of Broadly Cross-Reactive HIV-1-Neutralizing Human Monoclonal Antibodies	p. 143
Selection of Antibodies Able to Rapidly Enter Mammalian Cells	p. 155
Identification of the Specificity of Isolated Phage Display Single-Chain Antibodies Using Yeast Two-Hybrid Screens	p. 165
Generation of Bispecific and Tandem Diabodies	p. 177
Intrabody Expression in Eukaryotic Cells	p. 195
High-Level Periplasmic Expression and Purification of scFvs	p. 205
Expression of Single-Chain Fv Fragments in E. coli Cytoplasm	p. 215
High-Level Expression of a Phage Display-Derived scFv in Pichia pastoris	p. 225
Index	p. 237
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