Centrifugal Separations in Biotechnology

Description

Centrifugal Separations in Biotechnology, Second Edition, is the only book on the market devoted to centrifugal separation in biotechnology. Key topics covered include a full introduction to centrifugation, sedimentation and separation; detailed coverage of centrifuge types, including batch and semi-batch centrifuges, disk-stack and tubular decanter centrifuges; methods for increasing solids concentration; laboratory and pilot testing of centrifuges; selection and sizing centrifuges; scale-up of equipment, performance prediction and analysis of test results using numerical simulation. Centrifugal Separations in Biotechnology, Second Edition, provides guidance on troubleshooting and optimizing centrifuges, and then goes on to explore the commercial applications of centrifuges in biotechnology. It gives detailed process information and data to assist in the development of particular processes from existing systems. It is of value to professionals in the chemical, bioprocess, and biotech sectors, and all those concerned with bioseparation, bioprocessing, unit-operations and process engineering. Cover Centrifugal Separations in Biotechnology Copyright Contents In God, I trust Preface to Second Edition (2019) Preface to First Edition (2007) 1 Introduction 1.1 Introduction 1.1.1 Common Host Cells for Secreting Recombinant Protein 1.1.2 Platform for Protein Expression 1.1.3 Extracellular Protein 1.1.4 Intracellular Protein—Liquid 1.1.5 Intracellular Protein—Inclusion Body 1.2 Centrifugal Separation and Filtration 1.2.1 Sedimenting Centrifuge 1.2.2 Filtering Centrifuges 1.3 Pros and Cons of Filtration Versus Centrifugation 1.4 Generic Flow Sheet for Biopharmaceutical Process 1.5 Other Centrifugal Separations 1.6 Inputs and Outputs of Centrifuge 1.7 Separation Metrics 1.7.1 Protein Yield 1.7.2 Centrate Suspended Solids 1.7.3 Throughput Rate 1.7.4 Cell Viability 1.8 Text Organization 1.9 Summary References Problems 2 Principles of Centrifugal Sedimentation 2.1 Introduction 2.2 Nonintuitive Phenomena 2.2.1 Pressure Gradient in a Fluid Under Centrifugal Acceleration 2.2.2 Combined Centrifugal and Gravitational Accelerations 2.2.3 Coriolis Effect 2.3 Intuitive Phenomena 2.3.1 Centrifugal Acceleration 2.3.2 Fluid in a Centrifuge Bowl Not at Solid-Body Rotation 2.3.3 Regimes of Sedimentation 2.3.4 Stokes’ Law 2.3.5 Settling With Concentrated Solids 2.4 Process Functions 2.5 Summary References Problems 3 Batch and Semibatch Centrifuges 3.1 Spintube 3.2 Centrifugal Filter 3.3 Ultracentrifuges 3.3.1 Analytical Ultracentrifuge 3.3.2 Preparative Ultracentrifuge 3.3.3 Centrifugal Elutriation 3.4 Tubular Centrifuge 3.4.1 General Tubular Bowl Geometry 3.4.2 Ribs and Solids Scraper 3.4.3 Automatic Plunger Cake Discharge 3.5 Summary References Problems 4 Disk Centrifuge 4.1 Lamella/Inclined Plate Settler 4.1.1 Inclined Plate Settler Principle 4.1.2 Complications in Inclined Plate Settler 4.2 Disk-Stack centrifuge 4.2.1 General Disk Geometry 4.2.2 Disk Angle 4.2.3 Disk Spacing 4.2.4 Process Functions of Disk Centrifuge 4.2.5 Feed Solids 4.2.6 Manual Disk Centrifuge 4.2.6.1 Clarification of the Light Phase 4.2.6.2 Separation 4.2.7 Intermittent Discharge 4.2.7.1 Two Intermittent Discharge Designs 4.2.7.2 Angle of Cone for Discharge 4.2.7.3 Discharge Frequency 4.2.7.4 Concentrate Solids Discharge 4.2.7.5 Intermittent Discharge Interruption 4.2.8 Chamber Bowl 4.2.9 Continuous Concentrate Discharge 4.2.9.1 External Nozzle discharge 4.2.9.2 Small-Diameter Concentrate Discharge 4.2.9.3 Internal Vortex Nozzle Discharge 4.2.9.4 Applications of Different Concentrate Discharge Designs 4.2.9.5 External Nozzle Designs 4.2.10 Liquid Discharge 4.2.10.1 Centripetal Pump for Liquid Discharge 4.2.10.2 Hermetic Seal Design at Liquid discharge 4.2.11 Solution to Adverse Heating Effect 4.3 Feed Inlet and Accelerator 4.3.1 Introduction to Low Shear 4.3.2 Hydro-Hermetic Feed Design 4.3.3 Power Loss 4.3.4 Feed Acceleration Visual and Quantitative Testing 4.3.5 Improved Feed Accelerator 4.3.5.1 Improved Accelerator Without Smoothing Disk Section 4.3.5.2 Improved Accelerator With Smoothing Disk Section 4.4 Other Considerations 4.4.1 Materials of Construction 4.4.2 Clean-in-Place 4.4.3 Sterilization-in-Place 4.4.4 Containment 4.4.5 Surface Finish 4.4.6 Temperature Control 4.4.7 Water Requirements 4.4.8 Noise Level 4.4.9 Explosion Proof Design 4.5 Examples of Commercial Disk-Stack Centrifuge 4.6 Summary References Problems 5 Decanter Centrifuge 5.1 Solid Bowl or Decanter Centrifuge 5.2 Feed Rate 5.3 Pool Depth 5.4 Rotation Speed and G-Force 5.5 Differential Speed 5.6 Sedimentation Enhancement Using Chemical 5.7 Three-Phase Separation 5.8 Cake Conveyance 5.8.1 Dry Beach 5.8.2 Hydraulic Assist 5.9 Summary References Problems 6 Commercial Applications of Centrifugation in Biotechnology 6.1 Generic Flow Sheet of Biopharmaceutical 6.2 Mammalian Cell 6.3 Yeast Processing 6.4 Hormones Processing 6.5 Insulin Production 6.6 Biotech Separation of Inclusion Bodies 6.7 Vaccines Processing 6.7.1 Concentrated Cell-Based Product 6.7.2 Serum Product 6.8 Enzymes Processing 6.8.1 Extracellular Enzymes 6.8.2 Intracellular Enzymes 6.9 Probiotic Processing 6.10 Aquaculture 6.11 Alternative Meat 6.12 Baker Yeast Processing 6.13 Omega-3 From Microalgae 6.14 Ethanol Production 6.15 Other Biotech Processing 6.15.1 Recovery of Coagulation Factors From Blood Plasma 6.15.2 Tissue From Animal Cells 6.15.3 Laboratory Concentration and Buffer Exchange Using Centrifugal Filter 6.16 Summary References Problems 7 Concentrating Solids by Centrifugation 7.1 Introduction 7.2 Concentrating Underflow 7.3 Compaction 7.4 Expression or Percolation 7.5 Compaction Testing 7.6 Compaction Pressure 7.6.1 Test-Tube Compaction 7.6.2 Decanter Compaction 7.6.3 Considerations of Cake Compaction 7.7 Recommendations for Increasing Solid Concentration in Underflow 7.8 Summary References Problems 8 Laboratory and Pilot Testing 8.1 Process Objectives 8.2 Solid, Liquid, and Suspension Properties 8.2.1 Solids Properties 8.2.2 Mother Liquid Properties 8.2.3 Feed Slurry Properties 8.3 Bench-Scale Testing 8.3.1 Separability 8.3.2 Flocculant and Coagulant in Bench Tests 8.3.3 Test Variables 8.3.4 Material Balance 8.3.4.1 Material Balance Consideration for Bench Scale 8.3.5 Acceleration and Deceleration Time Duration 8.3.6 Settling Velocity 8.3.6.1 Apparatus for Visualizing Sedimentation Behavior 8.3.6.2 Sedimentation Behavior for Mondispersed Suspension 8.3.6.3 Sedimentation Behavior for Polydispersed Suspension 8.3.6.4 Sedimentation Behavior for Monodispersed Suspension With Hindered Settling 8.4 Centrifugal Filter Testing 8.4.1 Steady-State Membrane Centrifugal Filtration to Determine Protein Diffusivity and Solubility 8.4.2 Transient Membrane Centrifugal Filtration to Determine Protein Osmotic Pressure and Membrane Resistance 8.5 Pilot Testing 8.5.1 Material Balance Consideration for Pilot/Production Scale 8.5.1.1 Material Balance by Volume Fraction 8.5.1.2 Material Balance by Mass Fraction 8.5.2 Product (Protein) Yield 8.5.3 Pilot Test Factors 8.5.3.1 Monitored Variables in Pilot Tests 8.5.3.2 Metrics of Pilot Tests 8.5.3.3 Flocculant and Coagulant in Pilot Tests 8.6 Summary References Problems 9 Selection and Sizing of Centrifuges 9.1 Selection 9.1.1 Introduction 9.1.2 Tubular Centrifuge Selection 9.1.3 Disk Centrifuge Selection 9.1.4 Centrifuge Comparison 9.2 Centrifuge Sizing 9.2.1 Sizes and Rates 9.2.2 Dimensionless Le Number 9.2.3 Spintube (Bottle) Centrifuge 9.2.4 Sizing for Disk Centrifuge 9.2.4.1 Efficiency η in Le Number 9.2.5 Sizing for Tubular, Chamber, and Decanter Centrifuge 9.3 Feed Particle Size Distribution 9.4 Performance of Tubular Centrifuge 9.5 Summary References Further Reading Problems 10 Troubleshoot and Optimization 10.1 Troubleshooting 10.1.1 Timescale of Occurrence 10.1.2 Mechanical or Process Problem 10.1.3 Process Problems 10.1.3.1 High Centrate Turbidity 10.1.3.1.1 High Feed Solids Throughput Causing High Centrate Turbidity 10.1.3.1.2 Finer Feed Solids Causing High Centrate Turbidity 10.1.3.1.3 Concentrate Not Discharging Causing High Centrate Turbidity 10.1.3.1.4 Lower Process Temperature and Higher Viscosity Causing High Centrate Turbidity 10.1.3.2 Wet/High-Moisture Concentrate 10.1.4 Mechanical Problem 10.1.4.1 High Vibration 10.1.4.2 Other Mechanical Problems 10.2 Optimization 10.2.1 Separation Metrics 10.2.2 Monitored Variables 10.2.3 Controlled Variables 10.2.4 Simple Optimization Scheme 10.2.4.1 Optimizing Disk, Tubular, Decanter, and Chamber Bowl Centrifuge 10.2.4.2 Optimizing Centrifuge (Increase Viscosity μ) 10.3 Summary Problems 11 Visualization and Modeling of Flow and Separation in Tubular Centrifuge 11.1 Flow Visualization 11.2 Improved Moving Layer Flow Model 11.3 Effect of Velocity Profile 11.4 Effect of Friction Within the Flow Layer 11.5 Dimensionless Le Parameter 11.6 Quantitative Prediction 11.6.1 Total Solids Recovery in Cake 11.6.2 Total Solids Recovery in the Centrate 11.6.3 Particle Size Distribution of Supernatant/Overflow 11.6.4 Cumulative Size Recovery 11.7 Sedimentation Tests 11.7.1 Experiments on Sedimentation in Rotating Bowl Centrifuge 11.8 Summary References Problems 12 Disk-Stack Modeling 12.1 Disk Model 12.1.1 Continuum Phase 12.1.2 Dispersed Phase 12.2 Model Validation 12.3 Complications 12.4 Summary References Problems 13 Performance Projection of Centrifuges in Bioseparation 13.1 Disk Centrifuge 13.1.1 Baseline Case (400-mm Disk) 13.1.2 Effect of Fine Size Distribution (400-mm Disk) 13.1.3 Effect of G-Force (580-mm disk) 13.1.4 Efficiency η in Le Number (580-mm Disk) 13.1.5 Disk Centrifuge for Yeast Processing (500-mm Disk) 13.1.6 Disk Centrifuge for Inclusion Body Separation (260-mm Disk) 13.1.7 Enzymes (580-mm Disk) 13.2 Tubular Centrifuge 13.2.1 High-G Tubular (150- and 300-mm Tubular) 13.2.2 Low-G Tubular (150- and 300-mm Tubular) 13.3 Decanter 13.4 Spintube 13.5 Further Discussion on Numerical Simulations 13.6 Summary References Problems 14 Rotating Membrane in Bioseparation 14.1 Membrane 14.1.1 Osmotic Pressure Resistance 14.1.2 Gel Resistance 14.1.3 Membrane Fouling and Cake Formation 14.1.4 Two Scenarios on Rotation 14.2 Rotating Disk Membrane With Surface Parallel to the G-Force 14.2.1 Dimensionless Numbers 14.2.2 Governing Equations and Solutions 14.2.2.1 Model 14.2.2.2 Approximate Analytical Solution 14.2.2.2.1 Mass Boundary Layer 14.2.2.2.2 Membrane Flux 14.2.3 Gel Concentration 14.2.4 Determining Diffusivity 14.2.5 Parametric Effects 14.2.5.1 Effect of Reynolds Number 14.2.5.2 Schmidt Number Effect 14.2.5.3 Effect of Bulk/Feed Concentration 14.3 Rotating Membrane With Membrane Perpendicular to the G-Force 14.3.1 Spintube Equipped With Membrane Module—Centrifugal Filter 14.3.2 Model on Swinging Bucket Equipped With Ultrafiltration Membrane 14.3.3 Comparing Test Results With Predictions 14.4 Summary References Problems 15 Flocculation With Decanter Centrifuges 15.1 Introduction 15.1.1 Coagulation and Flocculation 15.1.2 Decanter Centrifuge 15.1.3 Problems 15.2 Monotonic Size Distribution Model 15.2.1 Moving Layer 15.2.2 Floc Model 15.2.3 Exponential Floc Size Distribution 15.2.4 Leung Number Calculation 15.2.5 Model Solution 15.3 Field Test 15.3.1 Two Decanter Tests at Wastewater Treatment 15.3.2 Determining In Situ Floc Size 15.3.2.1 Matching Data to Model Prediction 15.4 Prediction 15.5 Scale-Up 15.5.1 Le scale-Up 15.5.2 Sigma Scale-Up 15.5.3 G/g-Volume Scale-Up 15.5.4 Surface Area Scale-Up 15.6 Summary References Further Reading Problems 16 Case Studies of Monotonic and Unimodal Size Distribution Models 16.1 Introduction 16.2 Monotonic Model Equivalent for Disk-Stack and Tubular Centrifuges 16.3 Disk-Stack Centrifuge for Processing Protein From Mammalian Cell Culture 16.3.1 Low Cell Density Cell Culture 16.3.2 High Cell Density Cell Culture 16.3.3 Centrate Solids and Turbidity 16.4 Tubular Centrifuge for Separating E. coli Lysate 16.5 Tubular Centrifuge for Separating S. pneumoniae Flocculate 16.6 Unimodal Size Distribution Model 16.6.1 Unflocculated Suspension 16.6.2 Flocculation in Disk-Stack Centrifuge 16.7 Comparing the Solids Recovery Between Monotonic and Unimodal Size Distributions 16.8 Summary References Problems 17 Classifying Bimodal Particle Size Distribution and Case Study of Inclusion Body Classification 17.1 Introduction 17.2 Mammalian Cells With Cell Debris 17.3 Processing Hybridoma Cell Broth 17.4 Bimodal Size Model 17.5 Application of Bimodal Model on Hybridoma Cell Separation 17.6 Variation in Fine Fractions From Debris 17.7 Size of Whole Cells 17.7.1 Whole Cells Average Size 17.7.2 Size Range on Whole Cells 17.8 Effect of Smaller Size (The Debris) 17.9 Size Recoveries 17.9.1 Size Recovery of Small (S) Particles in Centrate 17.9.2 Size Recovery of Large (L) Particles in Centrate 17.9.3 Example on Classification 17.9.4 Smaller Size Fraction Further Apart From Larger Size Fraction in Bimodal Feed 17.9.5 Smaller Size Fraction Closer to the Larger Size Fraction in Bimodal Feed 17.10 Classification of Inclusion Bodies 17.10.1 Conventional Inclusion Bodies Processing 17.10.2 New Inclusion Bodies Processing 17.11 Centrifuges for Inclusion Bodies processing 17.11.1 Disk-Stack Centrifuge 17.11.2 Tubular Centrifuge 17.11.3 Spintube Centrifuge 17.12 Separation by Size and Density Difference 17.13 Summary References Problems 18 Integration of Unified Modeling With Practice in Centrifugal Separations 18.1 Introduction 18.2 Unified Modeling to Centrifugal Separation 18.3 Applications of the Unified Separation Models 18.3.1 Analysis of Test Data 18.3.2 Prediction/Forecast 18.3.3 Guiding Testing 18.3.4 Optimization 18.3.5 Troubleshooting 18.3.6 Scale-Up/Scale-Down 18.4 Integration of Unified Separation Models With Practice 18.5 Summary Appendix A: Nomenclature Subscripts Symbols Appendix B: Buckingham-π Analysis for Decanter and Tubular, Disk-Stack, and Spintube Centrifuges B1 Decanter and Tubular Centrifuge (Separation/Clarification) B2 Disk-Stack Centrifuge (Separation/Clarification) B3 Spintube Centrifuge (Separation/Clarification) Appendix C: Centrate or Concentrate Discharge Through Rotating Impeller Appendix D: Answers to Problems in Chapters 2–17 Chapter 2 Chapter 3 Chapter 4 Chapter 5 Chapter 6 Chapter 7 Chapter 8 Chapter 9 Chapter 10 Chapter 13 Chapter 14 Chapter 15 Solutions Chapter 16 Solutions Chapter 17 Solutions Index Back Cover