Aggregation of Therapeutic Proteins

"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)

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.

Preface xvii Contributors xxi 1. Fundamental Structures and Behaviors of Proteins 1 Jennifer S. Laurence and C. Russell Middaugh 1.1 The Problem of Protein Aggregation 1 1.2 Parallels to Protein Folding 11 1.3 Views of Protein Stability and Aggregation 12 1.4 Models of Aggregation 22 1.5 Models of Protein Folding 29 1.6 Influences of Chemical Alteration on Aggregation 40 1.7 Approaches to Predicting Aggregation 46 1.8 Conclusions 49 References 50 2. Protein Aggregation Pathways, Kinetics, and Thermodynamics 63 Yi Li and Christopher J. Roberts 2.1 Introduction 63 2.2 Native and Nonnative Aggregation Pathways 66 2.3 Thermodynamics of Reversible Self-Association 69 2.4 Aggregation Kinetics and Distinguishing Kinetic Pathways 75 2.5 Chemical Modifications 80 2.6 Effects of Cosolvents or Cosolutes 82 AppendixDerivation of „t32 for van der Waals (vdW) Mixture 94 Acknowledgments 97 References 97 3. Identification and Impact of Aggregation-Prone Regions in Proteins and Therapeutic Monoclonal Antibodies 103 Sandeep Kumar, Xiaoling Wang, and Satish K. Singh 3.1 Introduction 103 3.2 Energy Landscapes, Protein Folding, and Aggregation 105 3.3 Prediction of APRs in Proteins and Biotherapeutics 106 3.4 Conclusions and Future Directions 114 Acknowledgments 115 References 115 4. External Factors Affecting Protein Aggregation 119 Wei Wang, Ning Li, and Stan Speaker 4.1 Introduction 119 4.2 Protein Aggregation Pathways 120 4.3 Effects of Temperature 132 4.4 Effects of Solution Conditions and Composition on Protein Aggregation 136 4.5 Effects of Processing Steps on Protein Aggregation 164 4.6 Effects of Solid-State Condition and Composition on Protein Aggregation 174 4.7 Summary 177 Acknowledgment 178 References 178 5. Experimental Detection and Characterization of Protein Aggregates 205 Vikas K. Sharma and Devendra S. Kalonia 5.1 Introduction 205 5.2 Aggregate Classifi cation 206 5.3 Analytical Tools for the Characterization of Aggregates 212 5.4 Summary 246 References 247 6. Approaches to Control Protein Aggregation during Bulk Production 257 Linda O. Narhi, Yijia Jiang, Rohini Deshpande, Sohye Kang, and Joseph Shultz 6.1 Introduction 257 6.2 Candidate Selection 257 6.3 Protein Aggregation and Cell Culture 269 6.4 Protein Aggregation and Purifi cation 271 6.5 Summary 295 References 295 7. Protein Aggregation and Particle Formation: Effects of Formulation, Interfaces, and Drug Product Manufacturing Operations 301 Hanns-Christian Mahler, Stefan Fischer, Theodore W. Randolph, and John F. Carpenter 7.1 Introduction 301 7.2 Roles of Conformational and Colloidal Stability in Reducing Rates of Aggregation 302 7.3 Effects of Interfaces on Protein Aggregation 305 7.4 Critical Processing Steps during Drug Product Manufacturing of Biopharmaceuticals 310 7.5 Particles in Parenteral Products and Visible Inspection 316 7.6 Summary and Outlook 324 References 325 8. Approaches to Managing Protein Aggregation in Product Development 333 Wei Wang and Nicholas W. Warne 8.1 Introduction 333 8.2 Approaches in Formulation Development 334 8.3 Protection of Proteins in Various Processing Steps 345 8.4 Aggregation Control by Structural Modifi cations 351 8.5 Summary 353 References 354 9. Case Studies Involving Protein Aggregation 367 Rahul S. Rajan, Tiansheng Li, and Tsutomu Arakawa 9.1 Introduction 367 9.2 Case Study 1: Aggregation in the Liquid State: The Role of Osmolytes in Stabilizing KGF toward Aggregation 368 9.3 Case Study 2: Aggregation in the Liquid State: Heterogeneity and Non-Linearity in IgG2 Aggregation during Long-Term Storage 376 9.4 Case Study 3: Aggregation in the Frozen State: The Role of Excipient Crystallization 381 9.5 Case Studies 4 and 5: Aggregation in the Lyophilized State: Role of Residual Moisture and Mechanisms of Excipient Stabilization 385 9.6 Case Study 6: Protein Particulation Due to