Protein Engineering for Therapeutics, Part A, Volume 502

Series Page

Contributors

Preface

Volume in Series

Simplified Synthetic Antibody Libraries

1. Introduction

2. Materials

3. Methods

Generation of Dual-Variable-Domain Immunoglobulin Molecules for Dual-Specific Targeting

1. Introduction

2. Design of DVD-Ig™ Molecules

3. Cloning and Expression of DVD-Ig™ Molecules

4. Molecular Characterization of DVD-Ig™ Proteins

5. Functional Characterization of DVD-Ig™ Proteins

6. Concluding Remarks

Acknowledgments

Discovery of Internalizing Antibodies to Tumor Antigens from Phage Libraries

1. Introduction

2. Selection of Tumor Cell-Specific Internalizing scFv from Phage Antibody Libraries

3. Profiling the Specificity of Phage Antibodies on Tumor Cells Using Flow Cytometry

4. Identification of the Cognate Antigen Recognized by Tumor-Specific scFv by Using Mass Spectrometry

5. Selection of Antigen-Specific Phage Antibodies by Sequential Selection on Tumor Cells Followed by Selection on Yeast Cells Displaying Specific Tumor Antigens

Optimizing Properties of Antireceptor Antibodies Using Kinetic Computational Models and Experiments

1. Introduction: Valence, Affinity, and Avidity in Antibodies

2. Quantifying Antibody Cross-linking Strength Using “VFC” Models of Antibody Binding to Cell-Surface Receptors

3. Workflow for Quantifying Antibody Cross-linking Strength

4. Model Applications and Insights

Acknowledgments

Cell-Specific siRNA Delivery by Peptides and Antibodies

1. Introduction

2. Cationic Cell Permeable Peptides

3. Solid Biodegradable Polymer and Vesicular (Liposomal) Systems

Acknowledgments

Conjugation of Anticancer Drugs Through Endogenous Monoclonal Antibody Cysteine Residues

1. Antibody Reduction

2. Monitoring Antibody Reduction

3. Conjugation with Maleimido Drug-Linker

4. Purification Methods

5. Small-Scale Conjugations for Early ADC Discovery

6. Procedure for Solid-Phase Conjugation

7. Determination of Drug Loading

8. Determination of Unconjugated Drug Content

9. Conclusions

Building and Characterizing Antibody-Targeted Lipidic Nanotherapeutics

1. Introduction

2. Preparation of Immunoliposomes

3. Physicochemical Characterization of Immunoliposomes

4. In Vitro and In Vivo Characterization of Immunoliposomal Therapeutics

Cell-Targeting Fusion Constructs Containing Recombinant Gelonin

1. Introduction

2. Engineered Proteins Targeting Her2/neu

3. rGel/BLyS Targeting Malignant B-Cells

4. VEGF121/rGel Targeting Tumor Vasculature and Skeletal Metastases

Fusions of Elastin-Like Polypeptides to Pharmaceutical Proteins

1. Introduction

2. Protein ELP Fusion Design

3. Recursive Directional Ligation by Plasmid Reconstruction

4. Expression and Purification of Protein ELP Fusions

5. Physical Characterization of Protein ELP Fusions

6. Conclusions

Acknowledgment

Oral Enzyme Therapy for Celiac Sprue

1. Introduction

2. Lead Identification

3. Assay Development

4. Gram-Scale Production and Formulation

5. Lead Optimization for Next-Generation Proteases

6. Summary and Outlook

Rational Design and Evaluation of Mammalian Ribonuclease Cytotoxins

1. Introduction

2. Attributes of Cytotoxic ptRNases

3. Assays to Evaluate the Cytotoxicity of ptRNases

4. Prospectus

Acknowledgments

Engineering Reduced-Immunogenicity Enzymes for Amino Acid Depletion Therapy in Cancer

1. Introduction: Amino Acid Depletion as a Chemotherapeutic Strategy

2. Method I. Combinatorial Deimmunization

3. Method II. Engineering Therapeutically Relevant Substrate Specificities into a Human Enzyme

4. Method III. Improving the Kinetics and Stability of a Human Enzyme

5. Concluding Remarks

This volume of Methods in Enzymology looks at Protein Engineering for Therapeutics. The chapters provide an invaluable resource for academics, researchers and students alike. With an international board of authors, this volume is split into sections that cover subjects such as Antibodies, Protein conjugates, Peptides, Enzymes and Scaffolds

• Chapters provide an invaluable resource for academics, researchers and students alike
• Iinternational board of authors
• This volume is split into sections that cover subjects such as Antibodies, Protein conjugates, Peptides, Enzymes and Scaffolds

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists