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.

Preface. Contributors. 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 Infl uences of Chemical Alteration on Aggregation. 1.7 Approaches to Predicting Aggregation. 1.8 Conclusions. 2. Protein Aggregation Pathways, Kinetics, and Thermodynamics  ( Yi Li and Christopher J. Roberts ). 2.1 Introduction. 2.2 Native and Nonnative Aggregation Pathways. 2.3 Thermodynamics of Reversible Self–Association. 2.4 Aggregation Kinetics and Distinguishing Kinetic Pathways. 2.5 Chemical Modifi cations. 2.6 Effects of Cosolvents or Cosolutes. 3. Identifi cation 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 APRs in Proteins and Biotherapeutics. 3.4 Conclusions and Future Directions. 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 Condition and Composition on Protein Aggregation. 4.7 Summary. 5. Experimental Detection and Characterization of Protein Aggregates  ( Vikas K. Sharma and Devendra S. Kalonia ). 5.1 Introduction. 5.2 Aggregate Classifi cation. 5.3 Analytical Tools for the Characterization of Aggregates. 5.4 Summary. 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 Purifi cation. 6.5 Summary. 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. 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 Modifi cations. 8.5 Summary. 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 KGF toward 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 and 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. 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. 11. Regulatory Perspective on Aggregates as a 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. Index.