Phage Display in Biotechnology and Drug Discovery details the theories, principles, and methods impacting the field and demonstrates applications for peptide phage display, protein phage display, and the development of novel antibodies.

Features:

• Describes the basic principles of phage display and the methods and systems which have been developed for key applications
• Outlines applications for peptide phage display in the development of vaccines and high throughput screens, as well as the exploration of protein-protein interactions and enzyme substrate characteristics
• Details the use of phage display in the engineering of protein stability, the identification and mapping of protein-protein interactions, and the development of catalysts
• Provides broad coverage of the impact of phage display technology on the development of protein therapeutics
• Shows how phage display and antibody libraries have been used to develop and improve therapeutic production
• Presents expert opinions on future challenges in the field

Contents

Filamentous Bacteriophage Structure and Biology

• Taxonomy and Genetics
• Viral Gene Products
• Structure of the Virion
• Filamentous Bacteriophage Life Cycle
• Phage Library Diversity
• Biological Bottlenecks: Sources of Library Censorship
• Quantitative Diversity Estimation
• Improved Library Construction

Vectors and Modes of Display

• Most Display Vectors are Based on Filamentous Phage
• General Cloning Vectors Based on Filamentous Phage
• Classification of Filamentous Phage Display Systems
• Phage f1-The First Phage-Display Vector
• Low DNA Copy Number Display Vectors Based on Fd-Tet
• Diversity of Type 3 Vectors
• Type 8 Vectors: First Lessons
• Mosaic Display in Type nn Systems
• Mosaic Display in Phagemid Systems
• Vectors for C-Terminal Display
• Phage Proteins as Constraining Scaffolds

Methods for the Construction of Phage-Displayed Libraries

• Oligonucleotide-Directed Mutagenesis
• Random Mutagenesis
• Combinatorial Infection and Recombination
• DNA Shuffling

Selection and Screening Strategies

• General Considerations
• The Selection Process
• Selections Methods

Phage Library for Developing Antibody-Targeted Diagnostics and Vaccines

• Phage-Display Libraries as Tools for Epitope Discovery
• Diagnostics
• Phage Libraries for Epitope Mapping
• Phage Display Libraries for Vaccine
• Development
• Developing Immunogens From Peptide Leads

Exploring Protein-Protein Interactions Using Peptide Libraries Displayed on Phage

• Extracellular Protein-Protein Interactions
• Intracellular Protein-Protein Interactions

Substrate Phage Display

• The Concept of Substrate Phage Display
• Application of Substrate Phage Display to Cancer Research

Mapping Intracellular Protein Networks

• Domain-Mediated Interactions
• Nondomain Mediated Protein-Protein Interactions
• Software for Identifying Candidate Interacting Partners
• Analyzing Predicted Interactions
• Relevance to Biotechnology and Drug Discovery

High Throughput and High Content Screening Using Peptides

• Peptides as Enzyme Inhibitors
• Peptides as Conformational Probes

Engineering Protein Folding and Stability

• Protein Redesign and Design
• Early Combinatorial Studies Aimed at Repacking the Cores of Proteins
• Phage Display in Engineering Protein Stability
• A Worked Example: Repacking the Hydrophobic Core of Ubiquitin
• Studies that Build on the Original Methods

Identification of Natural Protein-Protein Interactions with CDNA Libraries

• Cloning Vectors
• Display of cDNA Libraries on Phage Surface
• Problems Associated with the Display of cDNA Libraries on Phage Surface
• Adaptability of Phage Display to High-Throughput Screening Technology

Mapping Protein Functional Epitopes

• Single Point Alanine Mutagenesis
• Combinatorial Site-Specific Mutagenesis
• Other Approaches to Phage-Displayed Functional Epitope Mapping

Selections for Enzymatic Catalysts

• Selection Methods

Antibody Humanization and Affinity Maturation Using Phage Display

• Humanization Using Phage Display
• In Vitro Affinity Maturation of Antibodies
• Emerging Approaches

Antibody Libraries from Immunized Repertoires

• Immune Antibody Library Construction
• Immune Antibody Library Selection

Naive Antibody Libraries from Natural Repertoires

• Construction of Naïve Libraries
• Applications of Naïve Libraries

Synthetic Antibody Libraries

• The Scripps Research Institute
• The Medical Research Council
• Morphosys
• Genentech

Index

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