Continuous processing in pharmaceutical manufacturing

Content: 
List of Contributors XVII 
Preface XXIII 
1 Proteins Separation and Purification by Expanded Bed Adsorption and Simulated Moving Bed Technology 1 Ping Li, Pedro Ferreira Gomes, Jose M. Loureiro, and Alirio E. Rodrigues 
1.1 Introduction 1 
1.2 Protein Capture by Expanded Bed Technology 3 
1.2.1 Adsorbent Materials 3 
1.2.2 Expanded Bed Adsorption/Desorption of Protein 10 
1.2.3 Modeling of the Expanded Bed 13 
1.3 Proteins Separation and Purification by Salt Gradient Ion Exchange SMB 15 
1.3.1 Adsorption Isotherms and Kinetics of BSA and Myoglobin on Ion Exchange Resins 16 
1.3.2 Salt Gradient Formation and Process Design for IE-SMB Chromatography 20 
1.3.3 Separation Region of Salt Gradient IE-SMB Chromatography 21 
1.3.4 Proteins Separation and Purification in Salt Gradient IE-SMB with Open Loop Configuration 24 
1.4 Conclusion 26 
References 27 
2 BioSMB Technology as an Enabler for a Fully Continuous Disposable Biomanufacturing Platform 35 Marc Bisschops 
2.1 Introduction 35 
2.2 Integrated Continuous Bioprocessing 36 
2.3 Multicolumn Chromatography 39 
2.4 BioSMB Technology 40 
2.5 Fully Disposable Continuous Processing 44 
2.6 Case Studies 46 
2.7 Regulatory Aspects 47 
2.8 Conclusions 50 
References 51 
3 Impact of Continuous Processing Techniques on Biologics Supply Chains 53 Aloke Das 
3.1 Introduction 53 
3.1.1 The Biologics Industry 53 
3.1.2 The Biologics Value Chain 54 
3.1.3 Downstream Purification Costs 54 
3.2 Chromatography Techniques Used in Downstream Purification of Biomolecules 55 
3.2.1 Need for Continuous Manufacturing in Downstream Purification 56 
3.2.2 The Multicolumn Countercurrent Solvent Gradient Purification Chromatography System 58 
3.3 Next-Generation Biologic Products     Bispecific Monoclonal Antibodies 59 
3.3.1 Major Biopharma Companies and Their Interest in Bispecific Mabs 59 
3.3.2 Challenges in Purification of Bispecific Monoclonal Antibodies 60 
3.4 Improving the Downstream Processing of Bispecific Mabs by Introduction of MCSGP in the Value Chain 61 
3.4.1 Advantages of Utilizing MCSGP Process in Bispecific Mabs Purification as Compared to Batch Chromatography 61 
3.4.2 Impact of MCSGP System on Biologic Supply Chains 62 
3.4.3 Impact on Patent Approval Structure of Biologic Drugs 62 
3.4.3.1 For a Manufacturer Who Already has a Biologic Drug in the Market 62 
3.4.3.2 For a Manufacturer Who is Developing a Biologic Drug 62 
3.4.4 Impact on Big Biopharma Companies 63 
3.4.5 Impact on the Chromatography Market 64 
3.4.6 Limitations of the MCSGP System 64 
3.5 Conclusion 64 
3.6 Further Research 65 
Acknowledgments 66 
3.A Appendix/Additional Information 66 
3.A.1 Regulatory Structure for Bispecific Monoclonal Antibodies 67 
3.A.1.1 Regulatory Compliance Comparison between US, EU, and Emerging Economies 67 
References 68 
4 Integrating Continuous and Single-Use Methods to Establish a New Downstream Processing Platform for Monoclonal Antibodies 71 Christopher Gillespie, Mikhail Kozlov, Michael Phillips, Ajish Potty, Romas Skudas, Matthew Stone, Alex Xenopoulos, Alison Dupont, Jad Jaber, and William Cataldo 
4.1 Introduction 71 
4.2 Harvest and Clarification 74 
4.2.1 The Challenge and Technology Selection 74 
4.2.1.1 Centrifugation 76 
4.2.1.2 Filtration 76 
4.2.1.3 Impurity Precipitation 77 
4.2.2 Summary 77 
4.3 Capture 78 
4.3.1 Background 78 
4.3.1.1 Protein A Chromatography 78 
4.3.2 Chromatographic Methods 79 
4.3.2.1 Slurried Bed Methods 79 
4.3.2.2 Continuous Chromatography 79 
4.3.3 Capture Case Studies 82 
4.3.3.1 Continuous Protein A Chromatography Capture Case Study 82 
4.3.3.2 Effect of Clarification Method on Protein A Performance 83 
4.4 Polishing 84 
4.4.1 Background 84 
4.4.2 Technology Selection Strategy 86 
4.4.3 Complete Flow-Through Polishing Case Study 87 
4.5 Cost of Goods Analysis 89 
4.5.1 Methodology 89 
4.5.2 Clarification 89 
4.5.3 Capture 90 
4.5.4 Polishing 91 
4.5.5 Overall 91 
4.6 Summary 92 
References 93 
5 Modeling of Protein Monomer/Aggregate Purification by Hydrophobic Interaction Chromatography: Application to Column Design and Process Optimization 97 Mark-Henry Kamga, Hae Woo Lee, Namjoon Kim, and Seongkyu Yoon 
5.1 Introduction 97 
5.2 Mathematical Model 99 
5.2.1 The Rate-Limiting Step in the HIC Process 99 
5.2.2 Dimensional Considerations 100 
5.2.2.1 Adsorption Capacity vs. Concentration of Vacant Sites (qmi vs. Cvi) 100 
5.2.2.2 Concentration of Protein Adsorbed on Resin (qi vs. Cia) 100 
5.2.3 Mathematical Model 101 
5.3 Experimentation 103 
5.3.1 Protein Solutions 103 
5.3.2 Determination of Adsorption and Desorption Kinetic Constants 104 
5.3.3 Column Chromatography 104 
5.4 Results and Discussion 105 
5.4.1 Kinetic Constants 105 
5.4.2 Protein Denaturation 107 
5.4.3 Model vs. Experimental Results 108 
5.4.4 Applications 109 
5.5 Conclusion 112 
Acknowledgments 112 
References 113 
6 Continuous Animal Cell Perfusion Processes: The First Step Toward Integrated Continuous Biomanufacturing 115 Leda R. Castilho 
6.1 Introduction 115 
6.2 The Basics of Perfusion Processes 116 
6.3 Cell Banking and Inoculum Development in the Context of Perfusion Processes 117 
6.4 Culture Conditions 120 
6.5 Cell Retention Devices 125 
6.5.1 Gravitational Settlers 126 
6.5.2 Centrifuges 130 
6.5.3 Hydrocyclones 131 
6.5.4 Acoustic (Ultrasonic) Separators 134 
6.5.5 Tangential Flow-Filtration 134 
6.5.6 ATF Systems 136 
6.5.7 Floating Membrane Devices 138 
6.5.8 Spin-Filters 138 
6.5.9 Rotating Cylindrical Filters (Vortex-Flow Filters or External Spin-Filters) 140 
6.5.10 Rotating Disc Filters (Controlled-Shear Filters) 141 
6.6 Integrated Perfusion   Purification Processes for Continuous Biomanufacturing 142 
6.7 Concluding Remarks 144 
References 145 
7 Perfusion Process Design in a 2D Rocking Single-Use Bioreactor 155 Nico M.G. Oosterhuis 
7.1 Introduction 155 
7.2 Production Costs 155 
7.3 Equipment Requirements for a Single-Use Perfusion Process 157 
7.4 Testing Results Single-Use Perfusion Process 159 
7.5 Simplified Seeding Process 161 
7.6 Future Outlook 163 
References 163 
8 Advances in the Application of Perfusion Technologies to Drosophila S2 Insects Cell Culture 165 Lars Poulsen and Willem A. de Jongh 
8.1 Introduction 165 
8.2 Case Study 1: Acoustic Separation 167 
8.2.1 The Perfusion Setup (BioSep) 167 
8.2.2 Results and Discussion 168 
8.2.2.1 Development 168 
8.2.2.2 Cell Count in the Bioreactor 168 
8.2.2.3 Effects of BioSep Settings on Cell Loss and Viability 169 
8.2.2.4 Controlling the Cell Concentration Through Bleed Rate Control 169 
8.2.2.5 Effect of Total Dilution Rate on Culture Viability 170 
8.2.2.6 Development of the Perfusion Rate Profile 170 
8.2.2.7 Initial Testing of Robustness of Upstream Process in 1.5 l Fermentations 170 
8.2.2.8 Scaling Up and Consistency in 4.5 l Fermentations 171 
8.2.2.9 Process Scale-Up 174 
8.2.3 Conclusions for Case Study 1 174 
8.3 Case Study 2: ATF-Based Cell Retention 176 
8.3.1 ATF Technology 176 
8.3.2 Methods 177 
8.3.3 Results 177 
8.3.3.1 Cell Counts Achieved Using Perfusion Technology 177 
8.3.3.2 Effect of Feed Strategy 178 
8.3.3.3 Yield Improvements Achieved Using Fed-Batch and Concentrated Perfusion 179 
8.3.3.4 Protein Stability 179 
8.3.4 Conclusions for Case Study 2 180 
8.4 Final Remarks 181 
Acknowledgments 181 
References 181 
9 Single-Use Systems Support Continuous Bioprocessing by Perfusion Culture 183 William G. Whitford 
9.1 Introduction 183 
9.2 Potential Advantages in Continuous Processing 187 
9.2.1 Improved Product Quality 187 
9.2.2 Ease in Process Development 188 
9.2.3 Improved Scalability 188 
9.2.4 Increased Profitability 189 
9.2.5 Sustainability 190 
9.3 Challenges in Adoption of Continuous Processing 190 
9.4 Continuous Biomanufacturing 194 
9.5 Single-Use Systems 196 
9.6 Hybrid Systems 202 
9.7 Perfusion Culture 203 
9.8 Single-Use in Continuous Biomanufacturing 205 
9.9 Roller Bottles 213 
9.10 Mechanically Agitated Suspension Reactors 213 
9.11 Hollow Fiber Media Exchange 214 
9.12 Packed Bed Bioreactors 215 
9.13 Hollow Fiber Perfusion Bioreactors 217 
9.14 Continuous Flow Centrifugation 218 
9.15 Acoustic Wave Separation 220 
9.16 Conclusion 222 
References 222 
10 Multicolumn Countercurrent Gradient Chromatography for the Purification of Biopharmaceuticals 227 Thomas Muller-Spath and Massimo Morbidelli 
10.1 Introduction to Multicolumn Countercurrent Chromatography 227 
10.2 Introduction to Multicolumn Simulated Moving Bed (SMB) Chromatography 230 
10.3 Capture Applications 232 
10.3.1 Introduction 232 
10.3.2 Process Principle 234 
10.3.3 Application Examples 236 
10.4 Polishing Applications 237 
10.4.1 Introduction 237 
10.4.2 Multicolumn Countercurrent Solvent Gradient Purification Principle 239 
10.4.3 Multicolumn Countercurrent Solvent Gradient Purification Process Design 242 
10.4.4 Multicolumn Countercurrent Solvent Gradient Purification Case Study 243 
10.5 Discovery and Development Applications 247 
10.6 Scale-Up of Multicolumn Countercurrent Chromatography 249 
10.7 Multicolumn Countercurrent Chromatography as Replacement for Batch Chromatography Unit Operations 249 
10.8 Multicolumn Countercurrent Chromatography in Continuous Manufacturing 251 
10.9 Process Analytical Tools for Multicolumn Countercurrent Processes 252 
References 253 
11 Monoclonal Antibody Continuous Processing Enabled by Single Use 255 Mark Brower, Ying Hou, and David Pollard 
11.1 Introduction 255 
11.1.1 Single-Use Revolution to Enable Process Intensification and Continuous Processing 256 
11.1.2 Principles of Continuous Multicolumn Chromatography for Biological Production (BioSMB) 260 
11.2 Continuous Downstream Processing for Monoclonal Antibodies Unit Operation Development 263 
11.2.1 Surge Vessels and Balancing Flows 265 
11.2.2 Primary Recovery: Centrifugation and Depth Filtration 266 
11.2.3 Bulk Purification: Continuous Multicolumn Chromatography     BioSMB Protein A Capture and Viral Inactivation 270 
11.2.3.1 Protein A Loading Zone Optimization 271 
11.2.3.2 Protein A Elution Zone Considerations 275 
11.2.3.3 Viral Inactivation 277 
11.2.4 Fine Purification: Flow-Through Anion Exchange Chromatography (AEX) 280 
11.2.4.1 Effects of Sample Flow Rate on AEX Membrane Chromatography 281 
11.2.4.2 Effect of Sample Loading Amount on AEX Membrane Chromatography 281 
11.2.4.3 Scaling-Up Membrane Chromatography for Continuous Processing 283 
11.2.5 Fine Purification: Continuous Multicolumn Chromatography     BioSMB Cation Exchange Chromatography 284 
11.2.5.1 Cation Exchange Loading Zone Optimization 284 
11.2.5.2 Cation Exchange Elution Zone Considerations 285 
11.2.6 Formulation: Continuous Ultrafiltration 287 
11.3 Pilot-Scale Demonstration of the Integrated Continuous Process 291 
11.4 Summary 293 
References 294 
12 Continuous Production of Bacteriophages 297 Ales   Podgornik, Nika Janez  , Franc Smrekar, and Matjaz   Peterka 
12.1 Bacteriophages 297 
12.1.1 Life Cycle 299 
12.1.2 Determination of Bacteriophage Properties 303 
12.2 Bacteriophage Cultivation 305 
12.2.1 Chemostat 306 
12.2.2 Cellstat 310 
12.2.3 Cellstat Productivity 314 
12.2.4 Bacteriophage Selection 322 
12.2.5 Technical Challenges 323 
12.3 Continuous Purification of Bacteriophages 325 
12.3.1 Centrifugation 326 
12.3.2 Precipitation and Flocculation 326 
12.3.3 Filtration 327 
12.3.4 Chromatographic and Other Adsorption Methods 328 
12.4 Conclusions 329 
References 329 
13 Very High Cell Density in Perfusion of CHO Cells by ATF, TFF, Wave Bioreactor, and/or CellTank Technologies     Impact of Cell Density and Applications 339 Veronique Chotteau, Ye Zhang, and Marie-Francoise Clincke 
13.1 Introduction 339 
13.2 Equipment 340 
13.3 Results and Discussion 342 
13.3.1 Perfusion Using ATF or TFF in Wave-Induced Bioreactor 342 
13.3.1.1 Cell Growth 342 
13.3.1.2 IgG Production 344 
13.3.2 Perfusion Using CellTank 346 
13.3.2.1 Cell Growth 346 
13.3.2.2 IgG Production 347 
13.3.3 Very High Cell Density 347 
13.3.4 Cryopreservation from Very High Cell Density Perfusion 350 
13.4 Conclusions 353 
Acknowledgments 354 
References 354 
14 Implementation of CQA (Critical Quality Attribute) Based Approach for Development of Biosimilars 357 Sanjeev K. Gupta 
14.1 Background 357 
14.2 Biosimilar Product Development 358 
14.3 Attributes/Parameters in Biopharmaceuticals 359 
14.3.1 Critical Quality Attributes 359 
14.3.2 Critical Process Parameters (CPP) 359 
14.3.3 The ICH Q8    Minimal Approach    to Pharmaceutical Development 359 
14.3.4 Quality-by-Design 360 
14.4 Quality Attributes and Biosimilars Development 361 
14.5 Quality, Safety, and Efficacy of Biosimilars 362 
14.6 Implementing CQA Approach for Biosimilar Development 364 
14.6.1 Identification of the CQA 364 
14.6.2 CQA-Based Clone Selection and Upstream Process Development 365 
14.6.3 Factors Affecting CQAs of the Biologics 366 
14.6.3.1 Expression Host and Recombinant Cell Line 366 
14.6.3.2 Process Related 367 
14.6.4 Protein Production Host and CQA 368 
14.6.4.1 Cell Line Changes and CQA 370 
14.6.4.2 Host Cell Line and Clone Selection Criteria 370 
14.6.5 Sequence Variants Identification by CQA 371 
14.6.6 Incomplete Processing of Signal Sequences and CQA 373 
14.6.7 Upstream Process Impact on Product Quality Attributes 374 
14.6.7.1 Bioreactor Optimization and Scale Up 374 
14.6.8 CQA in Purification and Formulation 379 
14.6.8.1 Downstream Processing of Biosimilars 379 
14.6.8.2 Downstream Processing and CQA 380 
14.6.9 CQAs in Formulation and Stability 381 
14.6.9.1 Formulation and Quality Attributes 381 
14.6.9.2 Stability and Quality Attributes 382 
14.7 Summary 382 
References 383 
15 Automated Single-Use Centrifugation Solution for Diverse Biomanufacturing Process 385 Sunil Mehta 
15.1 Introduction 385 
15.2 Separation by Centrifugation 385 
15.3 Separation by Filtration 386 
15.4 Downstream Process Challenges of High Cell Density Cultures 386 
15.5 Single-Use Centrifugation 387 
15.6 kSep Technology 387 
15.7 kSep System Configuration 390 
15.8 Low-Shear Process 392 
15.9 Perfusion 393 
15.10 Concentration, Media Replacement, and Harvest of Cells 395 
15.11 Continuous Harvest Clarification 397 
15.12 Separation of Cells from Microcarriers 398 
15.13 Summary 399 
References 399 
16 The Review of Flexible Production Platforms for the Future 401 Maik W. Jornitz 
16.1 Introduction 401 
16.2 Today   s Processing Technology Advances 402 
16.2.1 Single-Use Liquid Hold and Mixing Bags 402 
16.2.2 Filtration and Purification Technologies 406 
16.2.3 Product and Component Transfer 413 
16.2.4 Aseptic Fluid Connections and Disconnections 415 
16.2.5 Single-Use Final Filling Systems 417 
16.3 Todays Facility Designs 418 
16.3.1 Construction and Design Types 418 
16.3.2 Process Location and Flow 422 
16.4 Future Processing and Facility Requirements 424 
16.4.1 Upstream Technologies 424 
16.4.2 Downstream Technologies 426 
16.4.3 Single-Use Engineering and Design 427 
16.4.4 Facilities and Process Design 428 
References 431 
17 Evaluating the Economic and Operational Feasibility of Continuous Processes for Monoclonal Antibodies 433 Suzanne S. Farid, James Pollock, and Sa V. Ho 
17.1 Introduction 433 
17.2 Background on Continuous Processing 434 
17.2.1 Perfusion Culture 434 
17.2.2 Semicontinuous Chromatography 436 
17.3 Tool Description 438 
17.4 Case Study 1: Fed-batch Versus Perfusion Culture for Commercial mAb Production 440 
17.5 Case Study 2: Semicontinuous Affinity Chromatography for Clinical and Commercial Manufacture 446 
17.6 Case Study 3: Integrated Continuous Processing Flowsheets 450 
17.7 Conclusions 452 
Acknowledgments 452 
References 453 
18 Opportunities and Challenges for the Implementation of Continuous Processing in Biomanufacturing 457 Sadettin S. Ozturk 
18.1 Introduction 457 
18.2 A Brief History of Continuous Processing in Biomanufacturing 458 
18.3 Opportunities for Continuous Processing in Biomanufacturing 459 
18.3.1 Higher Process Yields 459 
18.3.2 Higher Process Efficiencies 461 
18.3.3 Compact and Flexible Facilities 461 
18.3.4 Stable and Consistent Production 462 
18.3.5 Better Product Quality 462 
18.4 Challenges for Implementing Continuous Processing in Biomanufacturing 462 
18.4.1 Process Complexity 463 
18.4.1.1 Cell Retention 463 
18.4.1.2 High Cell Density 466 
18.4.1.3 Longer Run Times 467 
18.4.2 Process Scalability in a Continuous Perfusion Process 470 
18.4.2.1 Scale and Capacity Limitations 470 
18.4.2.2 Process Scale-up 471 
18.4.3 Process Consistency and Control 473 
18.4.4 Process Characterization and Validation 474 
18.4.4.1 Complexity of a Scale-down Model for a Perfusion Process 474 
18.4.4.2 Process Optimization and Characterization for a Perfusion Process 475 
18.4.4.3 Process Validation 475 
18.5 Conclusions 476 
Acknowledgment 476 
References 476 
19 The Potential Impact of Continuous Processing on the Practice and Economics of Biopharmaceutical Manufacturing 479 L. Richard Stock, Marc Bisschops, and Thomas C. Ransohoff 
19.1 Introduction 479 
19.2 Background (Review of Status Quo     How We Make Biopharmaceutical Products Today) 480 
19.3 The Rationale for Continuous Processing 483 
19.4 The Obstacles for Implementation of Continuous Processing for Biopharmaceuticals 485 
19.5 The Potential Impact of Continuous Manufacturing on Process Economics 487 
19.6 The Potential Impact of Continuous Processing on Biopharmaceutical Manufacturing Practices 490 
19.7 Summary 492 
References 492 
Index 495