Aggregation Of Therapeutic Proteins

This book gives pharmaceutical scientists an up-to-date resource on protein aggregation and its consequences, and available methods to control or slow down the aggregation process. While significant progress has been made in the past decade, the current understanding of protein aggregation and its consequences is still immature. Prevention or even moderate inhibition of protein aggregation has been mostly experimental. The knowledge in this book can greatly help pharmaceutical scientists in the development of therapeutic proteins, and also instigate further scientific investigations in this area. This book fills such a need by providing an overview on the causes, consequences, characterization, and control of the aggregation of therapeutic proteins.

WEI WANG, PhD, is a Research Fellow at Pfizer Global Biologics. He is also Adjunct Professor in the School of Pharmacy and Health at the University of the Pacific in California and Guest Professor at Shandong University in China.
CHRISTOPHER J. ROBERTS, PhD, is Associate Professor of Chemical Engineering at the University of Delaware. Previously, he was a senior research scientist in pharmaceutical R & D at Pfizer.

"Among their topics are fundamental structures and behaviors of proteins; protein aggregation pathways, kinetics, and thermodynamics; the identification and impact of aggregate-prone regions in proteins and therapeutic monoclonal antibodies; external factors affecting protein aggregation; experimental detection and characterization of protein aggregates; approaches to controlling it during bulk production; effects of formulation interfaces, and drug product manufacturing operations on aggregation and particle formation; approaches to managing it in product development; aggregation and immunogenicity of therapeutic proteins; and a regulatory perspective on aggregates as a product quality attribute." (SciTech Book News, December 2010)

Chapter 1. Fundamental Structures and Behaviors of Proteins (Jennifer S. Laurence and C. Russell Middaugh).
1.1 The Problem of Protein Aggregation.
1.2 Parallels to Protein Folding.
1.3 Views of Protein Stability and Aggregation.
1.4 Models of Aggregation.
1.5 Models of Protein Folding.
1.6 Influences of Chemical Alteration on Aggregation.
1.7 Approaches to Predicting Aggregation.
1.8 Conclusions.
1.9 References.
Chapter 2. Protein Aggregation Pathways, Kinetics, and Thermodynamics (Yi Li and Christopher J. Roberts).
2.1 Introduction.
2.2 Native and Non-Native Aggregation Pathway(s).
2.3 Thermodynamics of Reversible Self-Association.
2.4 Aggregation Kinetics and Distinguishing Kinetic Pathways.
2.5 Chemical Modifications.
2.6 Effects of Cosolvents or Cosolutes.
2.7 Nomenclature.
2.8 Appendix - Derivation of Γ32 for a van der Waals Mixture.
2.9 Acknowledgements.
2.10 References.
Chapter 3. Identification and Impact of Aggregation Prone Regions in Proteins and Therapeutic Monoclonal Antibodies (Sandeep Kumar, Xiaoling Wang, and Satish K. Singh).
3.1 Introduction.
3.2 Energy Landscapes, Protein Folding and Aggregation.
3.3 Prediction of Aggregation Prone Regions in Proteins and Biotherapeutics.
3.4 Conclusions and Future Directions.
3.5 Acknowledgments .
3.6 Abbreviations.
3.7 References.
Chapter 4. External Factors Affecting Protein Aggregation (Wei Wang, Ning Li, and Stan Speaker).
4.1 Introduction.
4.2 Protein Aggregation Pathways.
4.3 Effects of Temperature.
4.4 Effects of Solution Conditions and Composition on Protein Aggregation.
4.5 Effects of Processing Steps on Protein Aggregation.
4.6 Effects of Solid-State Conditions and Composition on Protein Aggregation.
4.7 Summary.
4.8 Acknowledgements.
4.9 References.
Chapter 5. Experimental Detection and Characterization of Protein Aggregates (Vikas K. Sharma and Devendra S. Kalonia).
5.1 Introduction.
5.2 Aggregate Classification.
5.3 Analytical Tools for Characterization of Aggregates.
5.4 Summary.
5.5 References.
Chapter 6. Approaches to Control Protein Aggregation during Bulk Production (Linda O. Narhi, Yijia Jiang, Rohini Deshpande, Sohye Kang, and Joseph Shultz).
6.1 Introduction.
6.2 Candidate selection.
6.3 Protein aggregation and cell culture.
6.4 Protein aggregation and purification.
6.5 Summary.
6.6 References.
Chapter 7. Protein Aggregation and Particle Formation: Effects of Formulation, Interfaces and Drug Product Manufacturing Operations (Hanns-Christian Mahler, Stefan Fischer, Theodore W. Randolph, and John F. Carpenter).
7.1 Introduction.
7.2 Roles of Conformational and Colloidal Stability in Reducing Rates of Aggregation.
7.3 Effects of Interfaces on Protein Aggregation.
7.4 Critical processing steps during drug product manufacturing of biopharmaceuticals.
7.5 Particles in parenteral products and visible inspection.
7.6 Summary and Outlook.
7.7 References.
Chapter 8. Approaches to Managing Protein Aggregation in Product Development (Wei Wang and Nicholas W. Warne).
8.1 Introduction.
8.2 Approaches in Formulation Development.
8.3 Protection of Proteins in Various Processing Steps.
8.4 Aggregation Control by Structural Modifications.
8.5 Summary.
8.6 References.
Chapter 9. Case Studies Involving Protein Aggregation (Rahul S. Rajan, Tiansheng Li, and Tsutomu Arakawa).
9.1 Introduction.
9.2 Case study 1. Aggregation in the liquid state: the role of osmolytes in stabilizing Keratinocyte Growth Factor (KGF) towards aggregation.
9.3 Case Study 2: Aggregation in the liquid state: heterogeneity and non-linearity in IgG2 aggregation during long term storage.
9.4 Case Study 3: Aggregation in the frozen state: the role of excipient crystallization.
9.5. Case Studies 4&5: Aggregation in the lyophilized state: role of residual moisture and mechanisms of excipient stabilization.
9.6 Case study 6: Protein particulation due to nucleation by foreign material in fill/finish manufacturing operations.
9.7 Overall Discussion.
9.8 Acknowledgements.
9.9 References.
Chapter 10. Aggregation and Immunogenicity of Therapeutic Proteins (Vasco Filipe, Andrea Hawe, Huub Schellekens, and Wim Jiskoot).
10.1 Introduction.
10.2 Immunogenicity of therapeutic proteins.
10.3 Immune mechanisms related to protein immunogenicity.
10.4 Aggregates and Immunogenicity.
10.5 Conclusions.
10.6 References.
Chapter 11. Regulatory Perspective on Aggregates as Product Quality Attribute (Wendy C. Weinberg, Linan Ha, Susan L. Kirshner, and Daniela I. Verthelyi).
11.1 Introduction.
11.2 An overview of the regulatory process.
11.3 Product aggregates and safety concerns.
11.4 The assessment of aggregates: regulatory approaches to controlling product aggregation.
11.5 Future challenges.
11.6 Summary.
11.7 Acknowledgements.
11.8 References.