Front Cover

Biophysical Characterization of Proteins in Developing Biopharmaceuticals

Copyright

Contents

List of Contributors

About the Editors

Preface

List of Abbreviations and Symbols

Chapter 1 - The Complexity of Protein Structure and the Challenges it Poses in Developing Biopharmaceuticals

1.1 THE BASICS OF PROTEIN HIGHER-ORDER STRUCTURE (HOS)

1.2 THE SEARCH FOR HOW PROTEINS ATTAIN THEIR CORRECT HOS: THE PROTEIN FOLDING PROBLEM

1.3 SURPRISES IN THE WORLD OF PROTEIN FOLDING: INTRINSICALLY DISORDERED OR UNSTRUCTURED PROTEINS (AN APPARENT CHALLENGE TO THE ...

1.4 PROTEINS AND THE BIOPHARMACEUTICAL INDUSTRY: PROBLEMS AND CHALLENGES

1.5 CONCLUSION

References

Further Reading

Chapter 2 - Biophysical Characterization and Its Role in the Biopharmaceutical Industry

2.1 DRUG DEVELOPMENT PROCESS

2.2 PROTEIN DRUGS (BIOPHARMACEUTICALS)

2.3 THE ROLE OF BIOPHYSICAL CHARACTERIZATION IN BIOPHARMACEUTICAL DRUG DEVELOPMENT

2.4 THE CHALLENGES IN CONDUCTING BIOPHYSICAL MEASUREMENTS TO DETECT CHANGES IN A PROTEIN DRUG'S HOS

2.5 REGULATORY NEEDS AND CONSIDERATIONS

References

Further Reading

Chapter 3 - Biopharmaceutical Industry's Biophysical Toolbox

3.1 ATTRIBUTES OF A SINGLE BIOPHYSICAL TOOL TO CHARACTERIZE AND DETECT CHANGES IN THE HIGHER ORDER STRUCTURE OF A BIOPHARMACEUTICAL

3.2 STUDYING THE BIOPHYSICAL PROPERTIES OF A BIOPHARMACEUTICAL AS AN INDIRECT APPROACH FOR CHARACTERIZING CHANGES IN ITS HOS

3.3 GENERAL CONSIDERATIONS IN ANALYZING THE BIOPHYSICAL PROPERTIES OF BIOPHARMACEUTICALS

3.4 THE UTILITY OF USING STRESS TO MONITOR CHANGES IN THE HOS PROFILE OF A PROTEIN DRUG

3.5 PRESENT BIOPHYSICAL TOOLBOX

3.6 CONCLUSION

References

Further Reading

Chapter 4 - An Introduction and Hierarchical Organization of the Biophysical Tool in Section II

4.1 INTRODUCTION

4.2 THE STANDARD CLASS OF BIOPHYSICAL TOOLS USED IN THE BIOPHARMACEUTICAL INDUSTRY

4.3 THE ADVANCED CLASS OF BIOPHYSICAL TOOLS USED IN THE BIOPHARMACEUTICAL INDUSTRY

4.4 AN OVERVIEW OF SECTION II

References

Further Reading

Chapter 5 - The Value of UV, Fluorescence, and FTIR Spectroscopy in Biopharmaceutical Development

5.1 INTRODUCTION

5.2 THE ORIGINS OF ELECTRONIC ABSORPTION, FLUORESCENCE, AND FT-IR SPECTROSCOPY

5.3 CONFORMATIONAL ANALYSIS OF PROTEINS IN SOLUTION

5.4 OPTICAL SPECTROSCOPY AND PRODUCT COMPARABILITY

5.5 OPTICAL SPECTROSCOPY AND HIGH-THROUGHPUT METHODS

5.6 SOLID-STATE MEASUREMENTS

5.7 CONCLUSIONS

References

Chapter 6 - Circular Dichroism Spectroscopy for Protein Characterization: Biopharmaceutical Applications

6.1 INTRODUCTION

6.2 INSTRUMENTATION

6.3 DATA GENERATED

6.4 GUIDE TO COLLECTING GOOD DATA

6.5 DATA PROCESSING AND ANALYSES

6.6 ROLE IN THE RESEARCH INDUSTRY

6.7 TECHNOLOGY AVAILABILITY

6.8 FUTURE DEVELOPMENTS

Acknowledgments

References

Further Reading

Chapter 7 - Size-Exclusion Chromatograph (SEC) in Biopharmaceutical Process Development

7.1 INTRODUCTION

7.2 BASIC THEORY OF NORMAL OR IDEAL SEC

7.3 MAXIMIZING SEC SEPARATION BY ENHANCING THE USAGE OF PORE VOLUME AND PORE STRUCTURE

7.4 CHARACTERISTICS OF PORE STRUCTURE

7.5 NONIDEAL SEC CHROMATOGRAPHY

7.6 ASSESSING AND MAINTAINING AN OPTIMUM SEC CHROMATOGRAPHY METHOD

7.7 DETECTORS

7.8 MULTIDETECTOR SEC

7.9 AGGREGATION

7.10 TECHNOLOGY ADVANCES

7.11 CONCLUSION

References

Chapter 8 - Scattering Techniques for the Characterization of Biopharmaceuticals

8.1 INTRODUCTION

8.2 INTENSITY- AND TIME-DEPENDENT LIGHT SCATTERING

8.3 GENERAL COMMENT CONCERNING SLS AND DLS

8.4 THE “DUST PROBLEM” IN SLS AND DLS

8.5 X-RAY SCATTERING: CHARACTERIZATION OF PROTEINS IN SOLUTION USING SMALL-ANGLE X-RAY SCATTERING

References

Chapter 9 - Characterizing Biopharmaceuticals using Analytical Ultracentrifugation

9.1 INTRODUCTION

9.2 UNIQUE FEATURES OF THE ANALYTICAL ULTRACENTRIFUGE THAT MAKE IT DIFFERENT FROM OTHER CENTRIFUGES

9.3 THEORY

9.4 UTILITY OF AUC IN THE BIOPHARMACEUTICAL INDUSTRY

9.5 BOUNDARY SV-AUC

9.6 BAND SV-AUC

9.7 SEDIMENTATION EQUILIBRIUM, SE-AUC

9.8 DENSITY-GRADIENT SE-AUC

9.9 AUC DETECTORS

9.10 MISCELLANEOUS HELPFUL INFORMATION ABOUT CONDUCTING AUC EXPERIMENTS

9.11 CONCLUSION

References

Further Reading

Chapter 10 - Subvisible and Visible Particle Analysis in Biopharmaceutical Research and Development

10.1 INTRODUCTION

10.2 OVERVIEW OF ANALYTICAL METHODS

10.3 GENERAL RECOMMENDATIONS AND PITFALLS FOR PARTICLE ANALYSIS

10.4 OUTLOOK AND CONCLUSIONS

References

Chapter 11 - Differential Scanning Calorimetry in the Biopharmaceutical Sciences

11.1 BACKGROUND

11.2 DSC INSTRUMENTS

11.3 PRACTICAL CONSIDERATIONS FOR DSC USE

11.4 DATA ANALYSIS

11.5 APPLICATIONS OF SOLUTION DSC IN BIOPHARMACEUTICAL DISCOVERY AND DEVELOPMENT

11.6 APPLICATIONS OF SOLID-SAMPLE DSC IN BIOPHARMACEUTICAL DISCOVERY AND DEVELOPMENT

11.7 CONCLUSIONS

Acknowledgments

References

Chapter 12 - Biophysical Mass Spectrometry for Biopharmaceutical Process Development: Focus on Hydrogen/Deuterium Exchange

12.1 INTRODUCTION

12.2 SYNOPSIS OF THE TECHNIQUE

12.3 MECHANISM OF EXCHANGE

12.4 ADVANCES IN THE TECHNIQUE

12.5 COMMERCIALIZATION

12.6 APPLICATIONS IN THE BIOPHARMACEUTICAL INDUSTRY

12.7 FUTURE PERSPECTIVE

Acknowledgments

References

Chapter 13 - One- and Two-Dimensional NMR Techniques for Biopharmaceuticals

13.1 PHYSICAL BASIS OF THE TECHNIQUE

13.2 THE APPROPRIATE TECHNIQUE FOR A PARTICULAR PROBLEM

13.3 METHOD REQUIREMENTS AND PERFORMANCE

13.4 DATA PROCESSING (PROCEDURES)

13.5 ROLE IN RESEARCH VS PROCESS DEVELOPMENT

13.6 TECHNOLOGY UPDATE: RECENT AND FUTURE ADVANCES AND UNIQUE APPLICATIONS

References

Further Reading

Chapter 14 - Biophysical Characterization: An Integral Part of the “Totality of the Evidence” Concept

14.1 BIOPHARMACEUTICAL DEVELOPMENT

14.2 AN INTRODUCTION TO THE “TOTALITY OF THE EVIDENCE” AND ITS MORE GLOBAL MEANING IN DEVELOPING BIOPHARMACEUTICALS

14.3 BIOPHYSICAL CHARACTERIZATION IN DEVELOPING PROTEIN BIOPHARMACEUTICALS

14.4 BUILDING A BIOPHARMACEUTICAL'S BIOPHYSICAL FINGERPRINT

14.5 DETECTING SMALL DIFFERENCES IN BIOPHARMACEUTICALS VIA BIOPHYSICAL CHARACTERIZATION MEASUREMENTS

14.6 CONCLUSION

References

Index