Cryopreservation: Applications in Pharmaceuticals and Biotechnology (Drug Manufacturing Technology Series, V. 5)

Contents 4 Foreword 12 Author Biographies 14 1. Introduction 17 The Chapter Contents 21 2. Principles of Large-Scale Cryopreservation of Cells, Microorganisms, Protein Solutions, and Biological Products 26 Principles in the Freezing and Thawing of Cells 26 Scale-Up 35 Intracellular Ice Formation 37 Cell Membrane, Its Permeability and Freeze-Thaw Phenomena 38 Cryoconcentration Effects during Freezing and Chemical Reactions in the Liquid Phase 47 Osmotic Effects during Freezing and Thawing of Cells 50 Cooling and Warming Rates during Freezing and Thawing 51 Cell Concentration Effects during Freezing 54 Cell Damage by Mechanical Stress during Freezing 55 Cell Damage Caused by the Freeze-Thaw Cycles 57 Warming Rates and Thawing 61 Freezing of Protein and Peptide Solutions 62 Laboratory Equipment for Freezing 63 Storage Conditions 65 Ice Nucleation 66 Membrane and Cell Models in Freezing and Thawing 67 The Freezing Process: Physicochemical Phenomena 68 Particle Entrapment by an Advancing Freezing Front 68 Dendritic Crystal Growth 70 Dendritic Crystal Pattern and Freezing of Protein Solutions 86 Dendritic Crystal Growth and Cooling Rates in Cell Preservation 89 Cell Damage 92 Use of Cryoprotectants in Preservation of Cells and Proteins 92 Biopharmaceutical Formulations of Protein Drugs for Lyophilization and Freeze-Thaw Steps 110 Freezing of Cell Suspensions and Fermentation Broths 128 Effects of Sample Size and Shape 129 Controlled Cell Damage Using Freezing and Thawing 130 Estimate and Analysis of Freezing and Melting Processes 130 Estimate of Freezing Rates 130 Analytical and Approximate Methods 132 Numerical Methods 137 Slab, Cylinder, and Granule Freezing 140 Slab/Cube Freezing 140 Spherical Granule Freezing 141 Cylinder Freezing 153 Scale-Up of Freezing and Thawing of Cell Suspensions and Biological Solutions 153 Summary 157 References 166 3. Large-Scale Cryopreservation: Process Development for Freezing and Thawing of Large Volumes of Cell Suspensions, Protein Solutions, and Biological Products 195 Introduction 195 Process and System Development 199 Process and System Design Options 200 Evaluation of the Existing Processes and Systems 200 Case Study 3.1: Large-Scale Process and System Development for Freezing and Granulation of Cell Suspensions Using Liquefied Gases 223 A Brief Description 223 Principles of System Design and Operation 226 Discussion of Case Study 3.1 251 Case Study 3.2: Cryopreservation of Biological Liquid Products in Freeze-Thaw Vessels with Extended Heat Transfer Surfaces FINS of Novel Design 258 Advanced Design of Large-Scale Cryopreservation Vessels with Extended Heat Transfer Surfaces 258 System Design and Performance 272 Case Study 3.3: Cryopreservation of Biological Liquid Products in Bags in Contact with Plates with Extended Heat Transfer Surfaces FINS 274 Freezing of Biological Products in Bags Compressed between Plates with Extended Heat Transfer Surfaces 274 Thawing 284 Introduction 284 Thawing as a Part of the Freezing/Granulation Process 295 Thawing in Containers with Finned Internal Heat Transfer Surfaces Case Study 2 298 Thawing in Bags Using Plates with Extended Heat Transfer Surfaces Fins Case Study 3.3 302 Summary 304 Freezing and Granulation of Large Quantities of Solutions and Suspensions 306 Freezing of Large Volumes of Biological Solutions Using Extended Heat Transfer Surfaces 312 Thawing of Large Quantities of Solutions and Suspensions 313 Enhanced Freezing in Small Containers 314 Enhanced Thawing in Small Containers 316 References 317 4. Method Selection Considerations for Long-Term Preservation of Mammalian Cells and Microorganisms 328 Introduction 328 Chapter Focus 328 The Methods 329 Chapter Organization and Contents 333 Definitions 333 Management of Seed Stock Inventory:The Master Seed Bank MSB 335 Seed Bank Development 335 Characterization of the Seed Bank 336 Seed Bank Security 336 Validation 338 Benefits 338 Key Considerations for Method Selection 338 The Specimen and the Application 339 Equipment and Reagent Needs 340 Additives and Cryoprotectants 343 Regulatory Compliance 344 Safety Considerations 354 Preservation Procedures 362 General Comments 362 Special Hints for Successful Outcome 363 Evaluation of Method Performance 365 Part I: Procedures for Preservation of Mammalian Cells 368 Part II: Procedures for Preservation of Bacteria 369 Part III: Procedures for Preservation of Virus-Infected Cells 373 Part IV: Other Support Procedures 375 Summary/Conclusion 378 References 378 5. Cryopreservation of Mammalian Cell Cultures 383 Introduction 383 Cryopreservation Provides Insurance Against 383 Cryopreservation Provides Time 384 Cryopreservation Provides Enhancement of Cell Properties and a Selection Method 385 Suggested Practice to Maintain Consistent Cell Stocks 386 Brief Theory of Cryopreservation 386 Cryoprotectant Additives 387 Penetrating vs. Nonpenetrating 388 Requirements for Cryoprotectants 388 Concentration 389 Quality 389 Sterilization and Storage 389 Safety Issues 389 Liquid Nitrogen Hazard 389 Biohazardous Material 390 DMSO: A Powerful Solvent 391 Preparing Cells to Freeze 391 Words of Caution 391 Cell Condition before Freezing 391 Ampoules 392 Freezing Mixture Preparation 393 Dispensing 397 Ampoule Sealing 397 Configuration Variations 398 Concerns about Differentiation Induction by DMSO 400 Cooling 400 Rates 400 Methods 401 Recovery 403 Thawing 403 Open Ampoule 404 Dilute Cell Suspension 404 Cryoprotectant Removal 405 Seeding the Cells 406 Postfreeze Checks 406 Storage 409 Temperature vs. Expected Storage Time 409 Liquid Nitrogen: Vapor Phase vs. Liquid Phase 410 Equipment 411 Storage Practices 411 Summary 412 References 413 Suppliers 418 6. Freeze-Drying of Human Live Virus Vaccines 419 Introduction 419 Human Live Virus Vaccines LVVs 420 Instability of Live Virus Vaccines 423 Formulation of Live Virus Vaccines 424 Lyophilization of Live Virus Vaccines 425 Equipment 425 Process Overview 429 Special Process Considerations for LVV 430 Validation of Lyophilization Process 435 Recommendations for Continued Improvements 439 References 440 7. The Design and Use of Thermal Transport Containers 446 Introduction 446 Transport System Selection Criteria 447 Temperature Range Requirement 447 Refrigerant and Transport Method Options 448 Shipping Method 451 Internal and External Factors 453 Defining Insulated Container/Transporter and Refrigerant Options 454 Insulation Quality Considerations 454 Refrigerant Considerations 457 Shipping Dynamics 459 Insulated Container/Transporter Qualification 461 Examples of the Thermal Performance Range of Commercially Available Transport Systems 464 Test Methods for Evaluating Package Performance 470 Thermal Performance Testing—Design to Qualification 470 Distribution Test Methods 472 User's Responsibilities 473 Use of Temperature Monitors in Frozen Shipments 474 Common Problems in the Cold-Chain 475 Sources of Products and Testing Services for Shipment of Frozen Materials 478 References 480 Index 482 A 482 B 482 C 482 D 484 E 484 F 485 G 486 H 486 I 487 L 487 M 488 N 489 O 489 P 489 Q 490 R 490 S 490 T 491 U 492 V 492 W 492 Y 493 Z 493 This book covers the principles of cryopreservation as they relate the preservation of viable cells and cell materials being developed for biopharmaceutical applications. Topics include: the principles of freezing and thawing cells, physiochemical phenomena, process and system design options, method selection considerations, preservation procedures, cryoprotectant additives, freeze-drying human live virus vaccines, and transport system selection criteria. Contributions from well-known experts such as Steven S. Lee, Thomas C. Pringle, William H. Siegel, Richard Wisniewski, and Fangdong Yin make this the single most important study available.