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)

• Preface xviiContributors xxi1. Fundamental Structures and Behaviors of Proteins 1Jennifer S. Laurence and C. Russell Middaugh1.1 The Problem of Protein Aggregation 11.2 Parallels to Protein Folding 111.3 Views of Protein Stability and Aggregation 121.4 Models of Aggregation 221.5 Models of Protein Folding 291.6 Influences of Chemical Alteration on Aggregation 401.7 Approaches to Predicting Aggregation 461.8 Conclusions 492. Protein Aggregation Pathways, Kinetics, and Thermodynamics 63Yi Li and Christopher J. Roberts2.1 Introduction 632.2 Native and Nonnative Aggregation Pathways 662.3 Thermodynamics of Reversible Self-Association 692.4 Aggregation Kinetics and Distinguishing Kinetic Pathways 752.5 Chemical Modifications 802.6 Effects of Cosolvents or Cosolutes 823. Identification and Impact of Aggregation-Prone Regions in Proteins and Therapeutic Monoclonal Antibodies 103Sandeep Kumar, Xiaoling Wang, and Satish K. Singh3.1 Introduction 1033.2 Energy Landscapes, Protein Folding, and Aggregation 1053.3 Prediction of APRs in Proteins and Biotherapeutics 1063.4 Conclusions and Future Directions 1144. External Factors Affecting Protein Aggregation 119Wei Wang, Ning Li, and Stan Speaker4.1 Introduction 1194.2 Protein Aggregation Pathways 1204.3 Effects of Temperature 1324.4 Effects of Solution Conditions and Composition on Protein Aggregation 1364.5 Effects of Processing Steps on Protein Aggregation 1644.6 Effects of Solid-State Condition and Composition on Protein Aggregation 1744.7 Summary 1775. Experimental Detection and Characterization of Protein Aggregates 205Vikas K. Sharma and Devendra S. Kalonia5.1 Introduction 2055.2 Aggregate Classifi cation 2065.3 Analytical Tools for the Characterization of Aggregates 2125.4 Summary 2466. Approaches to Control Protein Aggregation during Bulk Production 257Linda O. Narhi, Yijia Jiang, Rohini Deshpande, Sohye Kang, and Joseph Shultz6.1 Introduction 2576.2 Candidate Selection 2576.3 Protein Aggregation and Cell Culture 2696.4 Protein Aggregation and Purifi cation 2716.5 Summary 2957. Protein Aggregation and Particle Formation: Effects of Formulation, Interfaces, and Drug Product Manufacturing Operations 301Hanns-Christian Mahler, Stefan Fischer, Theodore W. Randolph, and John F. Carpenter7.1 Introduction 3017.2 Roles of Conformational and Colloidal Stability in Reducing Rates of Aggregation 3027.3 Effects of Interfaces on Protein Aggregation 3057.4 Critical Processing Steps during Drug Product Manufacturing of Biopharmaceuticals 3107.5 Particles in Parenteral Products and Visible Inspection 3167.6 Summary and Outlook 3248. Approaches to Managing Protein Aggregation in Product Development 333Wei Wang and Nicholas W. Warne8.1 Introduction 3338.2 Approaches in Formulation Development 3348.3 Protection of Proteins in Various Processing Steps 3458.4 Aggregation Control by Structural Modifi cations 3518.5 Summary 3539. Case Studies Involving Protein Aggregation 367Rahul S. Rajan, Tiansheng Li, and Tsutomu Arakawa9.1 Introduction 3679.2 Case Study 1: Aggregation in the Liquid State: The Role of Osmolytes in Stabilizing KGF toward Aggregation 3689.3 Case Study 2: Aggregation in the Liquid State: Heterogeneity and Non-Linearity in IgG2 Aggregationduring Long-Term Storage 3769.4 Case Study 3: Aggregation in the Frozen State: The Role of Excipient Crystallization 3819.5 Case Studies 4 and 5: Aggregation in the Lyophilized State: Role of Residual Moisture and Mechanismsof Excipient Stabilization 3859.6 Case Study 6: Protein Particulation Due to Nucleation by Foreign Material in Fill/Finish Manufacturing Operations 3919.7 Overall Discussion 39410. Aggregation and Immunogenicity of Therapeutic Proteins 403Vasco Filipe, Andrea Hawe, Huub Schellekens, and Wim Jiskoot10.1 Introduction 40310.2 Immunogenicity of Therapeutic Proteins 40410.3 Immune Mechanisms Related to Protein Immunogenicity 40910.4 Aggregates and Immunogenicity 41510.5 Conclusions 42711. Regulatory Perspective on Aggregates as a Product Quality Attribute 435Wendy C. Weinberg, Linan Ha, Susan L. Kirshner, and Daniela I. Verthelyi11.1 Introduction 43511.2 An Overview of the Regulatory Process 43611.3 Product Aggregates and Safety Concerns 43811.4 The Assessment of Aggregates: Regulatory Approaches to Controlling Product Aggregation 44011.5 Future Challenges 44611.6 Summary 447Acknowledgments 447Disclaimer 447References 448Index 453

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)

• Preface xviiContributors xxi1. Fundamental Structures and Behaviors of Proteins 1Jennifer S. Laurence and C. Russell Middaugh1.1 The Problem of Protein Aggregation 11.2 Parallels to Protein Folding 111.3 Views of Protein Stability and Aggregation 121.4 Models of Aggregation 221.5 Models of Protein Folding 291.6 Influences of Chemical Alteration on Aggregation 401.7 Approaches to Predicting Aggregation 461.8 Conclusions 492. Protein Aggregation Pathways, Kinetics, and Thermodynamics 63Yi Li and Christopher J. Roberts2.1 Introduction 632.2 Native and Nonnative Aggregation Pathways 662.3 Thermodynamics of Reversible Self-Association 692.4 Aggregation Kinetics and Distinguishing Kinetic Pathways 752.5 Chemical Modifications 802.6 Effects of Cosolvents or Cosolutes 823. Identification and Impact of Aggregation-Prone Regions in Proteins and Therapeutic Monoclonal Antibodies 103Sandeep Kumar, Xiaoling Wang, and Satish K. Singh3.1 Introduction 1033.2 Energy Landscapes, Protein Folding, and Aggregation 1053.3 Prediction of APRs in Proteins and Biotherapeutics 1063.4 Conclusions and Future Directions 1144. External Factors Affecting Protein Aggregation 119Wei Wang, Ning Li, and Stan Speaker4.1 Introduction 1194.2 Protein Aggregation Pathways 1204.3 Effects of Temperature 1324.4 Effects of Solution Conditions and Composition on Protein Aggregation 1364.5 Effects of Processing Steps on Protein Aggregation 1644.6 Effects of Solid-State Condition and Composition on Protein Aggregation 1744.7 Summary 1775. Experimental Detection and Characterization of Protein Aggregates 205Vikas K. Sharma and Devendra S. Kalonia5.1 Introduction 2055.2 Aggregate Classifi cation 2065.3 Analytical Tools for the Characterization of Aggregates 2125.4 Summary 2466. Approaches to Control Protein Aggregation during Bulk Production 257Linda O. Narhi, Yijia Jiang, Rohini Deshpande, Sohye Kang, and Joseph Shultz6.1 Introduction 2576.2 Candidate Selection 2576.3 Protein Aggregation and Cell Culture 2696.4 Protein Aggregation and Purifi cation 2716.5 Summary 2957. Protein Aggregation and Particle Formation: Effects of Formulation, Interfaces, and Drug Product Manufacturing Operations 301Hanns-Christian Mahler, Stefan Fischer, Theodore W. Randolph, and John F. Carpenter7.1 Introduction 3017.2 Roles of Conformational and Colloidal Stability in Reducing Rates of Aggregation 3027.3 Effects of Interfaces on Protein Aggregation 3057.4 Critical Processing Steps during Drug Product Manufacturing of Biopharmaceuticals 3107.5 Particles in Parenteral Products and Visible Inspection 3167.6 Summary and Outlook 3248. Approaches to Managing Protein Aggregation in Product Development 333Wei Wang and Nicholas W. Warne8.1 Introduction 3338.2 Approaches in Formulation Development 3348.3 Protection of Proteins in Various Processing Steps 3458.4 Aggregation Control by Structural Modifi cations 3518.5 Summary 3539. Case Studies Involving Protein Aggregation 367Rahul S. Rajan, Tiansheng Li, and Tsutomu Arakawa9.1 Introduction 3679.2 Case Study 1: Aggregation in the Liquid State: The Role of Osmolytes in Stabilizing KGF toward Aggregation 3689.3 Case Study 2: Aggregation in the Liquid State: Heterogeneity and Non-Linearity in IgG2 Aggregationduring Long-Term Storage 3769.4 Case Study 3: Aggregation in the Frozen State: The Role of Excipient Crystallization 3819.5 Case Studies 4 and 5: Aggregation in the Lyophilized State: Role of Residual Moisture and Mechanismsof Excipient Stabilization 3859.6 Case Study 6: Protein Particulation Due to Nucleation by Foreign Material in Fill/Finish Manufacturing Operations 3919.7 Overall Discussion 39410. Aggregation and Immunogenicity of Therapeutic Proteins 403Vasco Filipe, Andrea Hawe, Huub Schellekens, and Wim Jiskoot10.1 Introduction 40310.2 Immunogenicity of Therapeutic Proteins 40410.3 Immune Mechanisms Related to Protein Immunogenicity 40910.4 Aggregates and Immunogenicity 41510.5 Conclusions 42711. Regulatory Perspective on Aggregates as a Product Quality Attribute 435Wendy C. Weinberg, Linan Ha, Susan L. Kirshner, and Daniela I. Verthelyi11.1 Introduction 43511.2 An Overview of the Regulatory Process 43611.3 Product Aggregates and Safety Concerns 43811.4 The Assessment of Aggregates: Regulatory Approaches to Controlling Product Aggregation 44011.5 Future Challenges 44611.6 Summary 447Acknowledgments 447Disclaimer 447References 448Index 453

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)

• Preface xviiContributors xxi1. Fundamental Structures and Behaviors of Proteins 1Jennifer S. Laurence and C. Russell Middaugh1.1 The Problem of Protein Aggregation 11.2 Parallels to Protein Folding 111.3 Views of Protein Stability and Aggregation 121.4 Models of Aggregation 221.5 Models of Protein Folding 291.6 Influences of Chemical Alteration on Aggregation 401.7 Approaches to Predicting Aggregation 461.8 Conclusions 492. Protein Aggregation Pathways, Kinetics, and Thermodynamics 63Yi Li and Christopher J. Roberts2.1 Introduction 632.2 Native and Nonnative Aggregation Pathways 662.3 Thermodynamics of Reversible Self-Association 692.4 Aggregation Kinetics and Distinguishing Kinetic Pathways 752.5 Chemical Modifications 802.6 Effects of Cosolvents or Cosolutes 823. Identification and Impact of Aggregation-Prone Regions in Proteins and Therapeutic Monoclonal Antibodies 103Sandeep Kumar, Xiaoling Wang, and Satish K. Singh3.1 Introduction 1033.2 Energy Landscapes, Protein Folding, and Aggregation 1053.3 Prediction of APRs in Proteins and Biotherapeutics 1063.4 Conclusions and Future Directions 1144. External Factors Affecting Protein Aggregation 119Wei Wang, Ning Li, and Stan Speaker4.1 Introduction 1194.2 Protein Aggregation Pathways 1204.3 Effects of Temperature 1324.4 Effects of Solution Conditions and Composition on Protein Aggregation 1364.5 Effects of Processing Steps on Protein Aggregation 1644.6 Effects of Solid-State Condition and Composition on Protein Aggregation 1744.7 Summary 1775. Experimental Detection and Characterization of Protein Aggregates 205Vikas K. Sharma and Devendra S. Kalonia5.1 Introduction 2055.2 Aggregate Classifi cation 2065.3 Analytical Tools for the Characterization of Aggregates 2125.4 Summary 2466. Approaches to Control Protein Aggregation during Bulk Production 257Linda O. Narhi, Yijia Jiang, Rohini Deshpande, Sohye Kang, and Joseph Shultz6.1 Introduction 2576.2 Candidate Selection 2576.3 Protein Aggregation and Cell Culture 2696.4 Protein Aggregation and Purifi cation 2716.5 Summary 2957. Protein Aggregation and Particle Formation: Effects of Formulation, Interfaces, and Drug Product Manufacturing Operations 301Hanns-Christian Mahler, Stefan Fischer, Theodore W. Randolph, and John F. Carpenter7.1 Introduction 3017.2 Roles of Conformational and Colloidal Stability in Reducing Rates of Aggregation 3027.3 Effects of Interfaces on Protein Aggregation 3057.4 Critical Processing Steps during Drug Product Manufacturing of Biopharmaceuticals 3107.5 Particles in Parenteral Products and Visible Inspection 3167.6 Summary and Outlook 3248. Approaches to Managing Protein Aggregation in Product Development 333Wei Wang and Nicholas W. Warne8.1 Introduction 3338.2 Approaches in Formulation Development 3348.3 Protection of Proteins in Various Processing Steps 3458.4 Aggregation Control by Structural Modifi cations 3518.5 Summary 3539. Case Studies Involving Protein Aggregation 367Rahul S. Rajan, Tiansheng Li, and Tsutomu Arakawa9.1 Introduction 3679.2 Case Study 1: Aggregation in the Liquid State: The Role of Osmolytes in Stabilizing KGF toward Aggregation 3689.3 Case Study 2: Aggregation in the Liquid State: Heterogeneity and Non-Linearity in IgG2 Aggregationduring Long-Term Storage 3769.4 Case Study 3: Aggregation in the Frozen State: The Role of Excipient Crystallization 3819.5 Case Studies 4 and 5: Aggregation in the Lyophilized State: Role of Residual Moisture and Mechanismsof Excipient Stabilization 3859.6 Case Study 6: Protein Particulation Due to Nucleation by Foreign Material in Fill/Finish Manufacturing Operations 3919.7 Overall Discussion 39410. Aggregation and Immunogenicity of Therapeutic Proteins 403Vasco Filipe, Andrea Hawe, Huub Schellekens, and Wim Jiskoot10.1 Introduction 40310.2 Immunogenicity of Therapeutic Proteins 40410.3 Immune Mechanisms Related to Protein Immunogenicity 40910.4 Aggregates and Immunogenicity 41510.5 Conclusions 42711. Regulatory Perspective on Aggregates as a Product Quality Attribute 435Wendy C. Weinberg, Linan Ha, Susan L. Kirshner, and Daniela I. Verthelyi11.1 Introduction 43511.2 An Overview of the Regulatory Process 43611.3 Product Aggregates and Safety Concerns 43811.4 The Assessment of Aggregates: Regulatory Approaches to Controlling Product Aggregation 44011.5 Future Challenges 44611.6 Summary 447Acknowledgments 447Disclaimer 447References 448Index 453