Protein engineering for therapeutics. Part B / edited by K. Dane Wittrup, Gregory L. Verdine.

Format:

Book

Contributor:

Wittrup, K. Dane, editor.

Verdine, Gregory L. (Gregory Lawrence), editor.

Series:

Methods in enzymology ; v. 503.

Methods in enzymology ; v. 503

Language:

English

Subjects (All):

Proteins--Therapeutic use.

Protein engineering.

Proteins.

Physical Description:

xliv, 348 pages, 6 unnumbered pages of plates : illustrations (some color) ; 23 cm.

Edition:

First edition.

Place of Publication:

San Diego, Calif. : Academic, 2012.

Contents:

Section IV Peptides 1

1 Stapled Peptides for Intracellular Drug Targets / Gregory L. Verdine Verdine, Gregory L., Gerard J. Hilinski Hilinski, Gerard J. 3

1 Introduction 4

2 All-Hydrocarbon Stapled α-Helical Peptides 6

3 The Design of Stapled Peptides 9

4 Stapled Peptide Synthesis 12

5 Olefin Metathesis 16

6 N-terminal and Internal Modifications 16

7 Cleavage of Stapled Peptides from the Solid Support 19

8 Stapled Peptide Purification 20

9 Biophysical Characterization 20

10 Cell Permeability 23

11 In Vitro Target Interaction and Activity Assays 26

12 In Vivo Efficacy 27

13 Strategies for Stapled Peptide Optimization 28

14 Summary 30

Acknowledgments 30

References 30

2 Mapping of Vascular ZIP Codes by Phage Display / Tambet Teesalu Teesalu, Tambet, Kazuki N. Sugahara Sugahara, Kazuki N., Erkki Ruoslahti Ruoslahti, Erkki 35

1 Introduction 36

2 Methods 41

3 Concluding Remarks and Perspectives 52

Acknowledgments 53

References 53

3 Engineering Cyclic Peptide Toxins / Richard J. Clark Clark, Richard J., David J. Craik Craik, David J. 57

1 Introduction 58

2 Peptide Design 59

3 Peptide Synthesis 60

4 Structural Analysis 66

5 Stability Assays 68

6 Special Considerations for Cyclic Peptides 69

Acknowledgments 71

References 71

4 Peptide Discovery Using Bacterial Display and Flow Cytometry / Jennifer A. Getz Getz, Jennifer A., Tobias D. Schoep Schoep, Tobias D., Patrick S. Daugherty Daugherty, Patrick S. 75

1 Introduction 76

2 Protocols 79

3 Conclusions 94

Acknowledgments 95

References 95

Section V Scaffolds 99

5 Designed Ankyrin Repeat Proteins (DARPins): From Research to Therapy / Rastislav Tamaskovic Tamaskovic, Rastislav, Manuel Simon Simon, Manuel, Nikolas Stefan Stefan, Nikolas, Martin Schwill Schwill, Martin, Andreas Plückthun Plückthun, Andreas 101

1 Introduction 102

2 Applications of DARPins 107

3 Protocols for DARPins in Biomedical Applications 112

Acknowledgment 129

References 129

6 Target-Binding Proteins Based on the 10th Human Fibronectin Type III Domain (¹⁰Fn3) / Shohei Koide Koide, Shohei, Akiko Koide Koide, Akiko, Dasa Lipovsek Lipovsek, Dasa 135

1 Introduction 136

2 Library Design 137

3 Choice of Selection Platform 139

4 Phage Display, mRNA Display, and Yeast-Surface Display of ¹⁰Fn3-Based Libraries 141

5 Production 151

6 Conclusion 153

Acknowledgments 154

References 154

7 Anticalins: Small Engineered Binding Proteins Based on the Lipocalin Scaffold / Michaela Gebauer Gebauer, Michaela, Arne Skerra Skerra, Arne 157

1 Introduction 158

2 Cloning and Expression of Lipocalins and Anticalins in E. coli 166

3 Construction of a Genetic Anticalin Library 170

4 Preparation and Selection of a Phage Display Library for Anticalins 172

5 Preparation and Selection of a Bacterial Surface Display Library for Anticalins 176

6 Colony Screening for Anticalins with Specific Target-Binding Activity 177

7 Screening for Anticalins with Specific Target-Binding Activity Using Microtiter Plate Expression in E. coli 179

8 Measuring Target Affinity of Anticalins in an ELISA 181

9 Measuring Target Affinity of Anticalins via Surface Plasmon Resonance 182

10 Application of Anticalins in Biochemical Research and Drug Development 184

References 186

8 T Cell Receptor Engineering / Jennifer D. Stone Stone, Jennifer D., Adam S. Chervin Chervin, Adam S., David H. Aggen Aggen, David H., David M. Kranz Kranz, David M. 189

1 Introduction 190

2 Stability and Affinity Engineering of T Cell Receptors by Yeast Surface Display 192

3 Affinity Engineering and Selection of T Cell Receptors by T Cell Display 200

4 Expression, Purification, and Applications of Soluble scTv Proteins 207

5 Recipes for Media and Buffers 215

Acknowledgments 218

References 219

9 Engineering Knottins as Novel Binding Agents / Sarah J. Moore Moore, Sarah J., Jennifer R. Cochran Cochran, Jennifer R. 223

1 Introduction 224

2 Knottins as Scaffolds for Engineering Molecular Recognition 225

3 Engineering Knottins by Yeast Surface Display 227

4 Knottin Library Construction 230

5 Screening Yeast-Displayed Knottin Libraries 234

6 Knottin production by chemical synthesis or recombinant expression 237

7 Cell binding assays 243

8 Summary 247

Acknowledgments 247

References 247

Section VI Pharmacokinetics 253

10 Practical Theoretic Guidance for the Design of Tumor-Targeting Agents / K. Dane Wittrup Wittrup, K. Dane, Greg M. Thurber Thurber, Greg M., Michael M. Schmidt Schmidt, Michael M., John J. Rhoden Rhoden, John J. 255

1 Introduction 256

2 What Molecular Size Is Best for Tumor Uptake? 256

3 Will Targeting Increase Nanoparticle Accumulation in a Tumor? 259

4 How Does Affinity Affect Biodistribution? 261

5 What Dose Is Necessary in Order to Overcome the "Binding Site Barrier"? 263

6 Conclusions 264

References 265

11 Reengineering Biopharmaceuticals for Targeted Delivery Across the Blood-Brain Barrier / William M. Pardridge Pardridge, William M., Ruben J. Boado Boado, Ruben J. 269

1 Introduction 270

2 Blood-Brain Barrier Receptor-Mediated Transport and Molecular Trojan Horses 271

3 Reengineering Recombinant Proteins for Targeted Brain Delivery 274

4 Genetic Engineering of Expression Plasmid DNA Encoding IgG Fusion Proteins 278

5 Pharmacokinetics and Brain Uptake of IgG Fusion Proteins 279

6 CNS Pharmacological Effects of IgG Fusion Proteins 284

7 Immune Response Against IgG Fusion Proteins 287

8 Summary 288

References 289

12 Engineering and Identifying Supercharged Proteins for Macromolecule Delivery into Mammalian Cells / David B. Thompson Thompson, David B., James J. Cronican Cronican, James J., David R. Liu Liu, David R. 293

1 Introduction 294

2 Methods 301

3 Conclusion 317

References 318.

Notes:

Includes bibliographical references and index.

ISBN:

9780123969620

012396962X

OCLC:

757491221