Description
Book SynopsisPromoting a continued and much-needed renaissance in biopharmaceutical manufacturing, this book covers the different strategies and assembles top-tier technology experts to address the challenges of antibody purification.
Updates existing topics and adds new ones that include purification of antibodies produced in novel production systems, novel separation technologies, novel antibody formats and alternative scaffolds, and strategies for ton-scale manufacturing
Presents new and updated discussions of different purification technologies, focusing on how they can address the capacity crunch in antibody purification
Emphasizes antibodies and innovative chromatography methods for processing
Table of ContentsPreface xxiii
List of Contributors xxvii
1 Downstream Processing of Monoclonal Antibodies: Current Practices and Future Opportunities 1
Brian Kelley
1.1 Introduction 1
1.2 A Brief History of Current Good Manufacturing Process mAb and Intravenous Immunoglobulin Purification 2
1.3 Current Approaches in Purification Process Development: Impact of Platform Processes 4
1.4 Typical Unit Operations and Processing Alternatives 7
1.5 VLS Processes: Ton‐Scale Production and Beyond 10
1.6 Process Validation 12
1.7 Product Life Cycle Management 13
1.8 Future Opportunities 16
1.9 Conclusions 18
Acknowledgments 19
References 19
2 The Development of Antibody Purification Technologies 23
John Curling
2.1 Introduction 23
2.2 Purification of Antibodies by Chromatography Before Protein A 25
2.3 Antibody Purification After 1975 28
2.4 Additional Technologies for Antibody Purification 31
2.5 Purification of mAbs Approved in North America and Europe 34
2.6 Current Antibody Process Technology Developments 40
Acknowledgments 45
References 46
3 Harvest and Recovery of Monoclonal Antibodies: Cell Removal and Clarification 55
Abhinav A. Shukla and Eric Suda
3.1 Introduction 55
3.2 Centrifugation 59
3.3 Microfiltration 62
3.4 Depth Filtration 67
3.5 Flocculation 70
3.6 Absolute Filtration 71
3.7 Expanded Bed Adsorption Chromatography 73
3.8 Harvesting in Single‐Use Manufacturing 74
3.9 Comparison of Harvest and Clarification Unit Operations 74
References 76
4 Next‐Generation Clarification Technologies for the Downstream Processing of Antibodies 81
Nripen Singh and Srinivas Chollangi
4.1 Introduction 81
4.2 Impurity Profiles in Cell Cultures 83
4.3 Precipitation 84
4.4 Affinity Precipitation 89
4.5 Flocculation 90
4.6 Toxicity of Flocculants and Precipitants and Their Residual Clearance 96
4.7 Depth Filtration 97
4.8 Considerations for the Implementation of New Clarification Technologies 102
4.9 Conclusions and Future Perspectives 103
Acknowledgments 104
References 104
5 Protein A‐Based Affinity Chromatography 113
Suresh Vunnum, Ganesh Vedantham and Brian Hubbard
5.1 Introduction 113
5.2 Properties of Protein A and Commercially Available Protein A Resins 114
5.3 Protein A Chromatography Step Development 118
5.4 Additional Considerations During Development and Scale‐Up 123
5.5 Virus Removal/Inactivation 127
5.6 Validation and Robustness 128
5.7 Conclusions 129
Acknowledgment 130
References 130
6 Purification of Human Monoclonal Antibodies: Non‐Protein A Strategies 135
Alahari Arunakumari and Jue Wang
6.1 Introduction 135
6.2 Integrated Process Design for Human Monoclonal Antibody Production 136
6.3 Purification Process Designs for HuMabs 136
6.4 Conclusions 149
Acknowledgments 151
References 152
7 Hydrophobic Interaction Chromatography for the Purification of Antibodies 155
Judith Vajda and Egbert Muller
7.1 Introduction 155
7.2 HIC With mAbs 156
7.3 HIC with Membrane Adsorbers 173
7.4 Future Perspectives 174
References 175
8 Purification of Monoclonal Antibodies by Mixed‐Mode Chromatography 181
Pete Gagnon
8.1 Introduction 181
8.2 A Brief History 182
8.3 Prerequisites for Industrial Implementation 183
8.4 Mechanisms, Screening, and Method Development 185
8.5 Capture Applications 192
8.6 Polishing Applications 193
8.7 Sequential Capture/Polishing Applications 193
8.8 Future Prospects 193
Acknowledgments 194
References 194
9 Advances in Technology and Process Development for Industrial‐Scale Monoclonal Antibody Purification 199
Nuno Fontes and Robert Van Reis
9.1 Introduction 199
9.2 Affinity Purification Platform 200
9.3 Advances in the Purification of mAbs by CEX Chromatography 201
9.4 High‐Performance Tangential Flow Filtration 209
9.5 A New Nonaffinity Platform 211
References 213
10 Alternatives to Packed‐Bed Chromatography for Antibody Extraction and Purification 215
Jorg Thommes, Richard M. Twyman and Uwe Gottschalk
10.1 Introduction 215
10.2 Increasing the Selectivity of Harvest Procedures: Flocculation and Filter Aids 216
10.3 Solutions for Antibody Extraction, Concentration, and Purification 218
10.4 Antibody Purification and Formulation Without Chromatography 220
10.5 Membrane Adsorbers 223
10.6 Conclusions 225
References 226
11 Process‐Scale Precipitation of Impurities in Mammalian Cell Culture Broth 233
Judy Glynn
11.1 Introduction 233
11.2 Precipitation of DNA and Protein—Other Applications 235
11.3 A Comprehensive Evaluation of Precipitants for the Removal of Impurities 236
11.4 Industrial‐Scale Precipitation 241
11.5 Cost of Goods Comparison 243
11.6 Summary 244
Acknowledgments 244
References 244
12 Charged Ultrafiltration and Microfiltration Membranes for Antibody Purification 247
Mark R. Etzel and Abhiram Arunkumar
12.1 Introduction 247
12.2 Charged UF Membranes 248
12.3 Concentration Polarization and Permeate Flux 248
12.4 Stagnant Film Model 249
12.5 Sieving Coefficient 250
12.6 Mass Transfer Coefficient 251
12.7 Mass Balance Models 251
12.8 Scale‐Up Strategies and the Constant Wall Concentration (Cw) Approach 253
12.9 Membrane Cascades 255
12.10 Protein Fractionation Using Charged UF Membranes 256
12.11 Case Study 257
12.12 Charged MF Membranes 259
12.13 Virus Clearance 260
12.14 Salt Tolerance 261
12.15 Conclusions 264
Acknowledgments 264
References 264
13 Disposable Prepacked‐Bed Chromatography for Downstream Purification: Form, Fit, Function, and Industry Adoption 269
Stephen K. Tingley
13.1 Introduction 269
13.2 Development‐Scale Prepacked Column Applications 271
13.3 Process‐Scale Prepacked Column Applications 275
13.4 Basic Technical Datasets 278
13.5 Independent Industry Assessments of “Fit for Purpose” 285
13.6 Case Study 1: Cation‐Exchange Polishing Chromatography 285
13.7 Case Study 2: Prepacked Columns for Pilot‐/Large‐Scale Bioprocessing 287
13.8 Prepacked Columns—Fit 292
13.9 The Economics of Prepacked Column Technologies 295
13.10 The Implementation of Disposable Prepacked Columns 297
13.11 Conclusions 300
References 301
14 Integrated Polishing Steps for Monoclonal Antibody Purification 303
Sanchayita Ghose, Mi Jin, Jia Liu, John Hickey and Steven Lee
14.1 Introduction 303
14.2 Polishing Steps for Antibody Purification 304
14.3 Integration of Polishing Steps 316
14.4 Conclusions 320
Acknowledgment 320
References 320
15 Orthogonal Virus Clearance Applications in Monoclonal Antibody Production 325
Joe X. Zhou
15.1 Introduction 325
15.2 Model Viruses and Virus Assays 326
15.3 Virus Clearance Strategies at Different Development Stages 328
15.4 Orthogonal Virus Clearance During mAb Production 328
15.5 Conclusions and Future Perspectives 338
Acknowledgments 339
References 339
16 Development of a Platform Process for the Purification of Therapeutic Monoclonal Antibodies 343
Yuling Li, Min Zhu, Haibin Luo and Justin R. Weaver
16.1 Introduction 343
16.2 Chromatography Steps in the Platform Process 345
16.3 Virus Inactivation 352
16.4 UF/DF Platform Considerations 352
16.5 Platform Development: Virus Filtration and Bulk Fill 354
16.6 Addressing Future Challenges in Downstream Processing 356
16.7 Representative Platform Processes 356
16.8 Developing a Virus Clearance Database Using a Platform Process 359
16.9 Summary 361
References 361
17 The Evolution of Platform Technologies for the Downstream Processing of Antibodies 365
Lee Allen
17.1 Introduction 365
17.2 The Definition of a Platform Purification Process 366
17.3 The Dominant Process Design 367
17.4 The Evolution of Unit Operations 372
17.5 Adapting the Platform Process for Product‐Specific Issues 382
17.6 Future Perspectives—Future Evolutionary Pathways 382
17.7 Concluding Remarks 383
Acknowledgments 384
References 384
18 Countercurrent Chromatography for the Purification of Monoclonal Antibodies, Bispecific Antibodies, and Antibody–Drug Conjugates 391
Thomas Muller‐Spath and Massimo Morbidelli
18.1 Introduction 391
18.2 Chromatography to Reduce Product Heterogeneity 392
18.3 Definition of Performance Parameters 394
18.4 Gradient Chromatography for Biomolecules 394
18.5 Continuous and Countercurrent Chromatography 395
18.6 Multicolumn Countercurrent Solvent Gradient Purification 397
18.7 Scalability of Multicolumn Countercurrent Chromatography 403
18.8 Online Process Monitoring for Multicolumn Countercurrent Chromatography 404
18.9 Outlook 405
References 405
19 The Evolution of Continuous Chromatography: From Bulk Chemicals to Biopharma 409
Marc Bisschops
19.1 Introduction 409
19.2 Continuous Chromatography in Traditional Process Industries 410
19.3 Continuous Chromatography in the Biopharmaceutical Industry 413
19.4 Advantages of Continuous Chromatography 420
19.5 Implementation Aspects of Continuous Chromatography 422
19.6 Regulatory Aspects 424
19.7 Conclusions 426
References 427
20 Accelerated Seamless Antibody Purification: Simplicity is Key 431
Benoit Mothes
20.1 Introduction 431
20.2 Accelerated Seamless Antibody Purification 432
20.3 Advantages of the ASAP Process 437
20.4 Scaling Up the ASAP Process 438
20.5 New Perspectives 440
20.6 Conclusion 442
Acknowledgments 442
Suggested Reading 443
21 Process Economic Drivers in Industrial Monoclonal Antibody Manufacture 445
Suzanne S. Farid
21.1 Introduction 445
21.2 Challenges When Striving for the Cost‐Effective Manufacture of mAbs 446
21.3 Cost Definitions and Benchmark Values 448
21.4 Economies of Scale 450
21.5 Overall Process Economic Drivers 453
21.6 DSP Drivers At High Titers 457
21.7 Process Economic Trade‐Offs for Downstream Process Bottlenecks 459
21.8 Summary and Outlook 461
References 462
22 Design and Optimization of Manufacturing 467
Andrew Sinclair
22.1 Introduction 467
22.2 Process Design and Optimization 468
22.3 Modeling Approaches 470
22.4 Process Modeling in Practice 481
22.5 Impact of the Process on the Facility 491
Acknowledgments 492
References 492
23 Smart Design for an Efficient Facility With a Validated Disposable System 495
Joe X. Zhou, Jason Li, Michael Cui and Haojun Chen
23.1 Design and Optimization of a Manufacturing Facility 495
23.2 Validation of a Disposable System 507
23.3 Conclusion 512
Acknowledgments 512
References 512
24 High‐Throughput Screening and Modeling Technologies for Process Development in Antibody Purification 515
Tobias Hahn, Thiemo Huuk and Jurgen Hubbuch
24.1 Introduction 515
24.2 Adsorption Isotherms 516
24.3 Batch Chromatography 519
24.4 Column Chromatography 524
References 532
25 Downstream Processing of Monoclonal Antibody Fragments 537
Mariangela Spitali
25.1 Introduction 537
25.2 Production of Antibody Fragments for Therapeutic Use 538
25.3 Downstream Processing 539
25.4 Improving the Pharmacological Characteristics of Antibody Fragments 552
25.5 Conclusions 553
Acknowledgments 555
References 555
26 Downstream Processing of Fc Fusion Proteins, Bispecific Antibodies, and Antibody–Drug Conjugates 559
Abhinav A. Shukla and Carnley L. Norman
26.1 Introduction 559
26.2 Biochemical Properties 562
26.3 Purification From Mammalian Expression Systems 576
26.4 Purification From Microbial Production Systems 585
26.5 Future Innovations 587
Acknowledgment 589
References 589
27 Manufacturing Concepts for Antibody–Drug Conjugates 595
Thomas Rohrer
27.1 Introduction 595
27.2 Targeting Components 596
27.3 Cytotoxic Drugs 600
27.4 Chemically Labile Linkers 602
27.5 General Process Overview 602
27.6 Facility Design and Supporting Technology 604
27.7 Single‐Use Equipment 607
27.8 Manufacturing ADCs 608
27.9 Analytical Support for ADC Manufacturing 609
27.10 Raw Materials Supply Chain 611
27.11 Conclusion 611
Acknowledgments 613
References 613
28 Purification of IgM and IgA 615
Charlotte Cabanne and Xavier Santarelli
28.1 Introduction 615
28.2 Purification of IgM 616
28.3 Purification of IgA 621
28.4 Conclusion 623
Acknowledgments 623
References 623
29 Purification of Monoclonal Antibodies From Plants 631
Zivko L. Nikolov, Jeffrey T. Regan, Lynn F. Dickey and Susan L. Woodard
29.1 Introduction 631
29.2 Antibody Production in Plants 632
29.3 Downstream Processing of Antibodies Produced in Plants 636
29.4 Purification of Plant‐Derived Antibodies Using Protein A Resins 641
29.5 Purification of Plant‐Derived Antibodies Using Non‐Protein A Media 642
29.6 Polishing Steps 643
29.7 Conclusions 645
Acknowledgment 645
References 645
30 Very‐Large‐Scale Production of Monoclonal Antibodies in Plants 655
Johannes F. Buyel, Richard M. Twyman and Rainer Fischer
30.1 Introduction 655
30.2 Process Schemes for mAb Production in Plants 656
30.3 Scalable Process Models 661
30.4 Process Adaptation for VLS Requirements 663
30.5 Translation into VLS Applications 666
References 667
31 Trends in Formulation and Drug Delivery for Antibodies 673
Hanns‐Christian Mahler and Roman Mathas
31.1 Introduction 673
31.2 Degradation Pathways 674
31.3 Physical Instability 674
31.4 Chemical Instability 676
31.5 How to Achieve Product Stability 678
31.6 Developability: Molecule Selection and Elimination of Degradation Hotspots 679
31.7 Stabilizing an Antibody in a Liquid Formulation 679
31.8 Stabilizing an Antibody by Drying 681
31.9 Choice of Adequate Primary Packaging 682
31.10 Minimizing Stress During Drug Product Processing 683
31.11 Implementation of a Formulation Strategy 685
31.12 Hot Topics 685
31.13 Summary 689
References 690
32 Antibody Purification: Drivers of Change 699
Narahari Pujar, Duncan Low and Rhona O’Leary
32.1 Introduction 699
32.2 The Changing Regulatory Environment—Pharmaceutical Manufacturing for the 21st Century 701
32.3 Technology Drivers—Advances and Innovations 707
32.4 Economic Drivers 708
32.5 Conclusions 711
Acknowledgment 712
References 713
Index 717
