Addressing a significant need by describing the science and process involved to develop biosimilars of monoclonal antibody (mAb) drugs, this book covers all aspects of biosimilar development: preclinical, clinical, regulatory, manufacturing.

Notes on Contributors xxv Preface xli 1 The History of Therapeutic Monoclonal Antibodies 1Regis Sodoyer 1.1 Summary 1 1.2 Introduction 1 1.3 New Markets for Old Antibodies, Old Markets for New Antibodies 2 1.4 Antibody Engineering: A New Approach to the Treatment of Disease 5 1.5 Fully Human Antibodies, What Else? 8 1.6 Antibody Design 17 1.7 Antibody Production 30 1.8 Recombinant Antibodies: No Limits… 37 Acknowledgments 37 References 37 2 Structure, Classification, and Naming of Therapeutic Monoclonal Antibodies 63Zhinan Xia 2.1 Summary 63 2.2 Introduction 64 2.3 Antibody Structure 65 2.4 Classification of Antibodies 71 2.5 IgG Subtype 73 2.6 Nomenclature of Therapeutic mAbs 73 2.7 List of Therapeutic mAbs on Market or in Review in the European Union and the United States 82 References 82 3 Mechanism of Action for Therapeutic Antibodies 85Yu Zhou and James D. Marks 3.1 Introduction 85 3.2 Blockade of Ligand–Receptor Interaction 86 3.3 Target Depletion via ADCC and CDC 94 3.4 Engaging Cytotoxic T Cell Through the Use of Bispecific Abs 95 3.5 Receptor Downregulation by Enhanced Internalization and Degradation 96 3.6 Targeted Drug Delivery 96 3.7 Summary 98 References 98 4 Therapeutic Monoclonal Antibodies and Their Targets 113Jose A. Figueroa, Camilo Pena, Leonardo Mirandola, Adair Reidy, J. Drew Payne, Nattamol Hosiriluck, Natallia Suvorava, Rakhshanda Layeequr Rahman, Adrienne R. Whitlow, Rashmi Verma, Everardo Cobos, and Maurizio Chiriva-Internati 4.1 Summary 113 4.2 Introduction 114 4.3 Monoclonal Antibody Therapies for Infectious Diseases 117 4.4 Monoclonal Antibody Therapies for Autoimmune Diseases 120 4.5 Therapeutic Monoclonal Antibodies Against Neoplastic Diseases 127 4.6 Conclusion 138 References 140 5 Antibody Posttranslational Modifications 155Roy Jefferis 5.1 Summary 155 5.2 Introduction 155 5.3 Overview of Co- and Posttranslational Modifications 157 5.4 Glycosylation 162 5.5 Glycation 172 5.6 IgG-Fab Glycosylation 179 5.7 The Influence of Expression Platform on CTM/PTMs and Unintended Physicochemical Changes 181 5.8 Human Antibody Isotypes Other than IgG 182 5.9 Conclusion 182 References 183 6 The Pharmacology, Pharmacokinetics, and Pharmacodynamics of Antibodies 201Ningning Xu, Meimei Liu, and Margaret Liu 6.1 Summary 201 6.2 Introduction 201 6.3 Pharmacology of Anticancer MAbs 202 6.4 Antibody Pharmacokinetics 204 6.5 Pharmacodynamics 208 6.6 Conclusions 211 References 211 7 Monoclonal Antibodies: Applications in Clinical Oncology 217Jeanene (“Gigi”) Robison 7.1 Summary 217 7.2 Introduction 217 7.3 Ado-trastuzumab Emtansine (Anti-HER2 Antibody Conjugated with Emtansine, Kadcyla®) 218 7.4 Alemtuzumab (Campath®, Campath-1H) 220 7.5 Bevacizumab (Avastin) 221 7.6 Brentuximab Vedotin (Anti-CD30 Antibody, Adcetris®) 225 7.7 Cetuximab (Anti-EGFR Antibody, Erbitux®) 227 7.8 Denosumab (Anti-RANKL Antibody, Xgeva™; Prolia™) 230 7.9 Eculizumab (Anti-C5 Antibody, Soliris®) 233 7.10 Ibritumomab Tiuxetan (Anti-CD20 Antibody, Zevalin®) 235 7.11 Ipilimumab (Anti-CTLA-4 Antibody, Yervoy®) 237 7.12 Obinutuzumab (Gazyva®) 238 7.13 Ofatumumab (Anti-CD20 Antibody, Arzerra®) 240 7.14 Panitumumab (Anti-EGFR Antibody, Vectibix™) 242 7.15 Pembrolizumab (Keytruda®) 244 7.16 Pertuzumab (Perjeta®) 246 7.17 Ramucirumab (Cyramza®) 248 7.18 Rituximab (Anti-CD20 Antibody, Rituxan) 250 7.19 Tositumomab and Iodine I-131 Tositumomab (Anti-CD20 Antibody, Bexxar®) 256 7.20 Trastuzumab (Anti-HER2 Antibody, Herceptin®) 258 References 262 8 Development of Biosimilar Rituximab and Clinical Experience 269Reena Nair 8.1 Summary 269 8.2 Introduction 270 8.3 Reditux Development Overview 271 8.4 Preclinical and Toxicology Studies 276 8.5 Clinical Evaluation 276 8.6 Conclusions 280 References 280 9 Monoclonal Antibodies for Infectious Diseases 283Steven J. Projan 9.1 Summary 283 9.2 Into the Future: Prophylaxis and Precision Medicine 283 9.3 Immune Therapy: A Noble Undertaking that Went to the Dogs 284 9.4 What’s Taking So Long? 285 9.5 Staphylococcus aureus: Still Public Enemy Number One? 285 9.6 Pseudomonas aeruginosa: The Bacterial Cockroach 286 9.7 Immune Evasion and Degree of Difficulty 287 9.8 Clostridium difficile: You Can’t Win for Losing 287 9.9 If Two Is Enough, Is Six Too Many? mAb Combos 288 9.10 Prophylaxis or Therapy? When You Come to a Fork in the Road, Take It 288 9.11 Influenza and Plan “B” 288 9.12 Safety: Human Enough for You? 288 9.13 Another Precinct Is Heard from Immunomodulatory Agents for the Treatment of Chronic Infections 289 9.14 Are We There Yet? Easy to Use, Fast Turnaround, Point-of-Care Diagnostics 289 9.15 Yeah but Aren’t These (Biologic) Drugs Going to Be Expensive? 290 References 290 10 Monoclonal Antibodies for Musculoskeletal, CNS, and Other Diseases 293Junming Yie and Tao Wu 10.1 Summary 293 10.2 Natalizumab (Tysabri®) 294 10.3 Eculizumab (Soliris®) 297 10.4 Ranibizumab (Lucentis®) 300 10.5 Denosumab (Prolia® and Xgeva®) 304 10.6 Antibody Therapies for Solid Organ Transplantation (Muromonab-CD3 (Orthoclone OKT3®), Basiliximab (Simulect®), and Daclizumab (Zenapax®)) 307 10.7 Conclusion 314 References 318 11 Manufacture of Recombinant Therapeutic Proteins Using Chinese Hamster Ovary Cells in Large-Scale Bioreactors: History, Methods, and Perspectives 327Florian M. Wurm and Maria de Jesus 11.1 Summary 327 11.2 Introduction 329 11.3 Process and Cells: The Quasi-species Concept Explains Individualized Development Needs 332 11.4 Choices for Manufacturing: Host Cells for Production and Suitable Selection Systems 335 11.5 Methods for Rapid Generation of High-Producing Cell Lines 337 11.6 Silencing: Stability of Expression, Facilitators for High-Level Productivity 339 11.7 High-Throughput Bioprocess Development 340 11.8 Disposable Bioreactors 342 11.9 Nonclonal Expression Technologies for Fast Production and Assessment of Expression Potential and Quality 343 11.10 Conclusions 345 Conflict of Interest 346 References 346 12 Process Development 355Samuel D. Stimple and David W. Wood 12.1 Summary 355 12.2 Introduction 355 12.3 Protein A and Protein G Batch Affinity Chromatography 356 12.4 Alternatives to Protein A 358 12.5 Disposables and Continuous Downstream Processing 361 12.6 Conclusion 373 References 374 13 Biosimilars and Biobetters: Impact on Biopharmaceutical Manufacturing and CMOs 381Ronald A. Rader 13.1 Summary 381 13.2 Introduction 382 13.3 The Biosimilar Pipeline 383 13.4 Developing Countries Will Continue to Prefer Cheaper Biogenerics 386 13.5 Biosimilar Candidates in the Pipeline 387 13.6 Biosimilar Development by Country/Region 387 13.7 Biosimilars Impact on Biopharmaceutical Markets and the Industry 389 13.8 Marketing Biosimilars Will Be a Challenge 391 13.9 Biosimilar Manufacturing Will Be State of the Art 391 13.10 Biosimilars Will Increase Demand for Product Quality and Transparency 392 13.11 CMOs Benefit from Biosimilars 393 13.12 Conclusions 394 References 395 14 Cell Line and Cell Culture Development for Biosimilar Antibody-Drug Manufacturing 397Jianguo Yang 14.1 Summary 397 14.2 Mammalian Cell Line Development 398 14.3 Cell Culture Process Development 406 14.4 Future Trends 418 References 419 15 Product Analysis of Biosimilar Antibodies 427Weidong Jiang, Scott Liu, and Ziyang Zhong 15.1 Summary 427 15.2 Introduction 428 15.3 Identity 428 15.4 Purity and Impurities 438 15.5 Stability 445 15.6 Quantity—Concentration Measurement 446 15.7 Biological Activity—Functional Bioassays 446 15.8 Efficacy and Safety: Animal Studies for Antibody-Drug Efficacy, PK/PD, and Toxicity 450 References 452 16 Bioanalytical Development 459Rafiq Islam 16.1 Summary 459 16.2 Introduction 459 16.3 Pharmacodynamics Characterization 460 16.4 Pharmacokinetic Assessment 465 16.5 Immunogenicity Assessment 472 16.6 Conclusion 474 References 475 17 Preclinical and Clinical Development of Biosimilar Antibodies 479João Eurico Fonseca and João Gonçalves 17.1 Summary 479 17.2 Introduction 480 17.3 Quality and Preclinical Development of Biosimilar Monoclonal Antibodies 481 17.4 Extrapolation of Indications 490 17.5 Clinical Development of Biosimilars of Monoclonal Antibodies 492 17.6 Ongoing Trials of Candidate Biosimilars of Monoclonal Antibodies 494 17.7 Conclusion 498 References 498 18 Regulatory Issues 505Clarinda Islam 18.1 Summary 505 18.2 Introduction 505 18.3 Existing Regulatory Pathways 506 18.4 Challenges 512 18.5 Conclusion 514 References 514 19 Legal Considerations 517K. Lance Anderson, Jennifer R. Moore Meline, and Jonathan D. Ball 19.1 Summary 517 19.2 Overview of the Biologics Price Competition and Innovation Act of 2009 (“BPCIA”) 519 19.3 Patent Litigation and the BPCIA 529 19.4 Patenting Your Biosimilar 541 19.5 Conclusion 543 Notes 544 20 ADCC Enhancement Technologies for Next-Generation Therapeutic Antibodies 549Cheng Liu and Su Yan 20.1 Summary 549 20.2 Introduction 549 20.3 Activation of ADCC Functions 550 20.4 ADCC Enhancement through Glycol-Engineering Technologies 552 20.5 Major ADCC Enhancement through Glycol-Engineering Technologies 553 20.6 ADCC Enhancement through Fc Mutagenesis 557 20.7 Major ADCC Enhancement Fc Mutagenesis Technologies 557 20.8 Conclusion 559 References 560 21 Antibody Half-Life: Engineering for Optimal Performance 565K. John Morrow, Jr. 21.1 Summary 565 21.2 Introduction 566 21.3 The IgG Molecule as a Therapeutic Entity 568 21.4 FcRn and Antibody Half-Life 569 21.5 Optimizing Antibody Fragments’ Half-Life 572 21.6 Albumin Fusions for Half-Life Extension 575 21.7 Mice as Models for Human Disease 577 21.8 Half-Life Engineering: Present and Future 578 21.9 A Bright Future for Biosimilars, Biobetters, and Improved Half-Life Modifications 583 References 585 22 Technologies for Antibody-Drug Conjugation 591Patrick G. Holder and David Rabuka 22.1 Summary 591 22.3 The Importance of Therapeutic Index 592 22.4 ADC Construction: Building from the Protein Out 593 22.5 Conjugation Sites and Heterogeneity 596 22.6 Installation of Conjugation Sites 597 22.7 Bioconjugation Reactions 602 22.8 Linking Antibodies and Payloads 613 22.9 Conclusion 623 References 623 Index 641

As the patents for biopharmaceuticals first marketed in the 1990s begin to expire, there is an opening for generic or non-proprietary versions of these agents to enter the market. The collective revenues of patent-expired biotech industry now total $10 billion annually, yet biosimilar generics have been slow to arise. It is a situation that has drawn much concern from generic and branded drug developers, patient groups, regulatory agencies and lawmakers. The 2012 release of FDA’s guidance on biosimilar drug development further stresses the need for a technical yet practical guide to biosimilar drug development. Arising from this landscape, Biosimilars of Monoclonal Antibodies: A Practical Guide to Manufacturing and Preclinical and Clinical Development gives pharmaceutical and biotech scientists and researchers a clear resource to understand the scientific principles and challenges involved in biosimilar drug development. The book discusses the scientific background including history, classification, and biological activities – including background knowledge unique to monoclonal antibody (mAb) drugs and essential profiling characteristics that regulations require. Contributing authors from the frontlines of biosimilar development address the processes and issues involved with manufacturing biosimilar mAbs, like cell line development, process development, large-scale cell culture of mammalian cells, and final product analysis. A valuable for all those –  from beginners to experts – with an interest in biosimilar drug development of monoclonal antibodies, Biosimilars of Monoclonal Antibodies: • Covers all aspects of biosimilar development: preclinical, clinical, regulatory, manufacturing• Introduces key topics of bioanalytical development, preclinical and clinical validation of biosimilarity, regulatory issues, and legal considerations concerning approval and commercialization• Leads readers to think beyond biosimilars by examining new technologies and strategies for improving biosimilar mAbs• Includes a review of FDA-approved mAb drugs as a quick reference to facts and useful information• Features flow charts, tables, and figures that clearly illustrate processes and makes the book comprehensible and accessible