Designed for undergraduates, graduate students, and industry practitioners, Bioseparations Science and Engineering fills a critical need in the field. Current, comprehensive, and concise, it covers bioseparations unit operations in greater depth than other texts on this topic. In each of the chapters, the authors use a consistent method of explaining unit operations, starting with a qualitative description noting the significance and general application of the unit operation. They then illustrate the scientific application of the operation, develop the required mathematical theory, and finally, describe the applications of the theory in engineering practice, with an emphasis on design and scaleup. Unique to this text is a chapter dedicated to bioseparations process design and economics, in which a process simulator, SuperPro® Designer®, is used to analyze and evaluate the production of three important biological products. Other unique features include basic information about bioproducts and engineering analysis and a chapter with bioseparations laboratory exercises. Bioseparations Science and Engineering is ideal for students and professionals alike. • Incorporates numerous example problems within the chapters • Offers extensive sets of problems at the end of chapters • Includes basic information about bioproducts • Provides thorough coverage of analytical methods for bioproducts • Uses the simulation software SuperPro® Designer® to illustrate the analysis and evaluation of the production of citric acid, recombinant human insulin, and monoclonal antibodies • Includes laboratory exercises that support text material • Supplemented by a website with new problems and examples and links to useful databases and manufacturers of bioseparations equipment and supplies Contents: All chapters (except Chapter 12) end with the following sections: Summary, Nomenclature, Problems, and References. Preface 1. Introduction to Bioproducts and Bioseparations 2. Analytical Methods 3. Cell Lysis and Flocculation 4. Filtration 5. Sedimentation 6. Extraction 7. Liquid Chromatography and Adsorption 8. Precipitation 9. Crystallization 10. Drying 11. Bioprocess Design 12. Laboratory Exercises in Bioseparations Appendix: Table of Units and Constants Index Ancillary material: • Solutions CD (019516153X) • Companion website: www.biosep.ou.edu

Preface All chapters (except Chapter 12) end with the following sections: Summary, Nomenclature, Problems, and References. Preface Chapter 1. Introduction to Bioproducts and Bioseparations 1.1. Instructional Objectives 1.2. Broad Classification of Bioproducts 1.3. Small Biomolecules 1.3.1. Primary Metabolites 1.3.2. Secondary Metabolites 1.3.3. Summary of Small Biomolecules 1.4. Macromolecules: Proteins 1.4.1. Primary Structure 1.4.2. Secondary Structure 1.4.3. Tertiary Structure Example 1.1: Effect of a Reducing Agent on Protein Structure and Mobility 1.4.4. Quaternary Structure 1.4.5. Prosthetic Groups and Hybrid Molecules 1.4.6. Functions and Commercial Uses of Proteins 1.4.7. Stability of Proteins 1.4.8. Recombinant Protein Expression 1.5. Macromolecules: Nucleic Acids and Oligonucleotides 1.6. Macromolecules: Polysaccharides 1.7. Particulate Products 1.8. Introduction to Bioseparations: Engineering Analysis 1.8.1. Stages of Downstream Processing Example 1.2: Initial Selection of Purification Steps 1.8.2. Basic Principles of Engineering Analysis 1.8.3. Process and Product Quality 1.8.4. Criteria for Process Development 1.9. The Route to Market 1.9.1. The Chemical and Applications Range of the Bioproduct 1.9.2. Documentation of Pharmaceutical Bioproducts 1.9.3. GLP and cGMP 1.9.4. Formulation Chapter 2. Analytical Methods 2.1. Instructional Objectives 2.2. Specifications 2.3. Assay Attributes 2.3.1. Precision 2.3.2. Accuracy 2.3.3. Specificity 2.3.4. Linearity, Limit of Detection, and Limit of Quantitation 2.3.5. Range 2.3.6. Robustness 2.4. Analysis of Biological Activity 2.4.1. Animal Model Assays 2.4.2. Cell-Line-Derived Bioassays 2.4.3. In Vitro Biochemical Assays Example 2.1: Coupled Enzyme Assay for Alcohol Oxidase 2.5. Analysis of Purity 2.5.1. Electrophoretic Analysis Example 2.2: Estimation of the Maximum Temperature in an Electrophoresis Gel 2.5.2. High Performance Liquid Chromatography (HPLC) 2.5.3. Mass Spectrometry 2.5.4. Coupling of HPLC with Mass Spectrometry 2.5.5. UV Absorbance Example 2.3: Determination of Molar Absorptivity 2.5.6. CHNO/Amino Acid Analysis (AAA) Example 2.4: Calculations Based on CHNO Analysis 2.5.7. Protein Assays 2.5.8. Enzyme-Linked Immunosorbent Assay 2.5.9. Gas Chromatography 2.5.10. DNA Hybridization 2.5.11. ICP/MS (AA) 2.5.12. Dry Weight 2.6. Microbiology Assays 2.6.1. Sterility 2.6.2. Bioburden 2.6.3. Endotoxin 2.6.4. Virus and Phage Chapter 3. Cell Lysis and Flocculation 3.1. Instructional Objectives 3.2. Some Elements of Cell Structure 3.2.1. Prokaryotic Cells 3.2.2. Eukaryotic Cells 3.3. Cell Lysis 3.3.1. Osmotic and Chemical Cell Lysis 3.3.2. Mechanical Methods of Lysis 3.4. Flocculation 3.4.1. The Electric Double Layer Example 3.1: Dependence of the Debye Radius on the Type of Electrolyte 3.4.2. Forces between Particles and Flocculation by Electrolytes Example 3.2: Sensitivity of Critical Flocculation Concentration to Temperature and Counter-Ion Charge Number 3.4.3. The Schulze-Hardy Rule 3.4.4. Flocculation Rate 3.4.5. Polymeric Flocculants Chapter 4. Filtration 4.1. Instructional Objectives 4.2. Filtration Principles 4.2.1. Conventional Filtration Example 4.1: Batch Filtration 4.2.2. Crossflow Filtration Example 4.2: Concentration Polarization in Ultrafiltration 4.3. Filter Media and Equipment 4.3.1. Conventional Filtration 4.3.2. Crossflow Filtration 4.4. Membrane Fouling 4.5. Scaleup and Design of Filtration Systems 4.5.1. Conventional Filtration Example 4.3: Rotary Vacuum Filtration Example 4.4: Washing of a Rotary Vacuum Filter Cake 4.5.2. Crossflow Filtration Example 4.5: Diafiltration Mode in Crossflow Filtration Chapter 5. Sedimentation 5.1. Instructional Objectives 5.2. Sedimentation Principles 5.2.1. Equation of Motion 5.2.2. Sensitivities 5.3. Methods and Coefficients 5.3.1. Equilibrium Sedimentation 5.3.2. Sedimentation Coefficient Example 5.1: Application of the Sedimentation Coefficient 5.3.3. Equivalent Time Example 5.2: Scaleup Based on Equivalent Time 5.3.4. Sigma Analysis 5.4. Production Centrifuges: Comparison and Engineering Analysis 5.4.1. Tubular Bowl Centrifuge Example 5.3: Complete Recovery of Bacterial Cells in a Tubular Bowl Centrifuge 5.4.2. Disk Centrifuge 5.5. Ultracentrifugation 5.5.1. Determination of Molecular Weight 5.6. Flocculation and Sedimentation 5.7. Sedimentation at Low Accelerations 5.7.1. Diffusion, Brownian Motion 5.7.2. Isothermal Settling 5.7.3. Convective Motion and Peclet Analysis 5.7.4. Inclined Sedimentation 5.7.5. Field-Flow Fractionation 5.8. Centrifugal Elutriation Chapter 6. Extraction 6.1. Instructional Objectives 6.2. Extraction Principles 6.2.1. Phase Separation and Partitioning Equilibria 6.2.2. Countercurrent Stage Calculations Example 6.1: Separation of a Bioproduct and an Impurity by Countercurrent Extraction Example 6.2: Effect of Solvent Rate in Countercurrent Staged Extraction of an Antibiotic 6.3. Scaleup and Design of Extractors 6.3.1. Reciprocating-Plate Extraction Columns Example 6.3: Scaleup of a Reciprocating-Plate Extraction Column 6.3.2. Centrifugal Extractors Chapter 7. Liquid Chromatography and Adsorption 7.1. Instructional Objectives 7.2. Adsorption Equilibrium 7.3. Adsorption Column Dynamics 7.3.1. Fixed-Bed Adsorption Example 7.1: Determination of the Mass Transfer Coefficient from Adsorption Breakthrough Curves 7.3.2. Agitated-Bed Adsorption 7.4. Chromatography Column Dynamics 7.4.1. Plate Models 7.4.2. Chromatography Column Mass Balance with Negligible Dispersion Example 7.2: Chromatographic Separation of Two Solutes Example 7.3: Calculation of the Shock Wave Velocity for a Non-Linear Isotherm Example 7.4: Calculation of the Elution Profile 7.4.3. Dispersion Effects in Chromatography 7.4.4. Gradients and Modifiers Example 7.5: Equilibrium for a Protein Anion in the Presence of a Chloride Ion 7.5. Adsorbent Types 7.5.1. Silica Based Resins 7.5.2. Polymer Based Resins 7.5.3. Ion Exchange Resins 7.5.4. Reversed Phase Chromatography 7.5.5. Hydrophobic Interaction Chromatography 7.5.6. Affinity Chromatography 7.5.7. Immobilized Metal Affinity Chromatography 7.5.8. Size Exclusion Chromatography 7.6. Particle Size and Pressure Drop in Fixed Beds 7.7. Equipment 7.7.1. Columns 7.7.2. Chromatography Column Packing Procedures 7.7.3. Detectors 7.7.4. Chromatography System Fluidics 7.8. Scaleup 7.8.1. Adsorption Example 7.6: Scaleup of the Fixed-Bed Adsorption of a Pharmaceutical Product 7.8.2. Chromatography Example 7.7: Scaleup of a Protein Chromatography Example 7.8: Scaleup of a Protein Chromatography Using Standard Column Sizes Example 7.9: Scaleup of Elution Buffer Volumes in Protein Chromatography Example 7.10: Consideration of Pressure Drop in Column Scaling Chapter 8. Precipitation 8.1. Instructional Objectives 8.2. Protein Solubility 8.2.1. Structure and Size 8.2.2. Charge 8.2.3. Solvent Example 8.1: Salting-Out of a Protein with Ammonium Sulfate. 8.3. Precipitation Formation Phenomena 8.3.1. Initial Mixing 8.3.2. Nucleation 8.3.3. Growth Governed by Diffusion Example 8.2: Calculation of Concentration of Nuclei in a Protein Precipitation Example 8.3: Diffusion-Limited Growth of Particles 8.3.4. Growth Governed by Fluid Motion Example 8.4: Growth of Particles Limited by Fluid Motion 8.3.5. Precipitate Breakage 8.3.6. Precipitate Aging 8.4. Particle Size Distribution in a Continuous Flow Stirred Tank Reactor Example 8.5: Dependence of Population Density on Particle Size and Residence Time in a CSTR 8.5. Methods of Precipitation 8.6. Design of Precipitation Systems Chapter 9. Crystallization 9.1. Instructional Objectives 9.2. Crystallization Principles 9.2.1. Crystals 9.2.2. Nucleation 9.2.3. Crystal Growth 9.2.4. Crystallization Kinetics from Batch Experiments 9.3. Batch Crystallizers 9.3.1. Analysis of Dilution Batch Crystallization Example 9.1: Batch Crystallization with Constant Rate of Change of Diluent Concentration 9.4. Process Crystallization of Proteins 9.5. Crystallizer Scaleup and Design 9.5.1. Experimental Crystallization Studies as a Basis for Scaleup 9.5.2. Scaleup and Design Calculations Example 9.2: Scaleup of Crystallization Based on Constant Power per Volume Chapter 10. Drying 10.1. Instructional Objectives 10.2. Drying Principles 10.2.1. Water in Biological Solids and in Gases Example 10.1: Drying of Antibiotic Crystals 10.2.2. Heat and Mass Transfer Example 10.2: Conductive Drying of Wet Solids in a Tray Example 10.3: Mass Flux during the Constant Rate Drying Period in Convective Drying Example 10.4: Time to Dry Nonporous Solids by Convective Drying 10.3. Dryer Description and Operation 10.3.1. Vacuum-Shelf Dryers 10.3.2. Batch Vacuum Rotary Dryers 10.3.3. Freeze Dryers 10.3.4. Spray Dryers 10.4. Scaleup and Design of Drying Systems 10.4.1. Vacuum-Shelf Dryers 10.4.2. Batch Vacuum Rotary Dryers 10.4.3. Freeze Dryers 10.4.4. Spray Dryers Example 10.5: Sizing of a Spray Dryer Chapter 11. Bioprocess Design 11.1. Instructional Objectives 11.2. Definitions and Background 11.3. Synthesis of Bioseparation Processes 11.3.1. Primary Recovery Stages 11.3.2. Intermediate Recovery Stages 11.3.3. Final Purification Stages 11.3.4. Pairing of Unit Operations in Process Synthesis 11.4. Process Analysis 11.4.1. Spreadsheets 11.4.2. Process Simulators 11.4.3. Using a Biochemical Process Simulator 11.5. Process Economics 11.5.1. Capital Cost Estimation 11.5.2. Operating Cost Estimation 11.5.3. Profitability Analysis 11.6. Illustrative Examples 11.6.1. Citric Acid Production 11.6.2. Human Insulin Production 11.6.3. Therapeutic Monoclonal Antibody Production Chapter 12. Laboratory Exercises in Bioseparations 12.1. Flocculant Screening 12.1.1. Background 12.1.2. Objectives 12.1.3. Procedure 12.1.4. Report 12.1.5. Some Notes and Precautions 12.2. Crossflow Filtration 12.2.1. Background 12.2.2. Objectives 12.2.3. Procedure 12.2.4. Report 12.3. Centrifugation of Flocculated and Unflocculated Particulates 12.3.1. Background 12.3.2. Objectives 12.3.3. Procedure 12.3.4. Report 12.4. Aqueous Two-Phase Extraction 12.4.1. Physical Measurements 12.4.2. Procedure 12.4.3. Calculations and Report 12.4.4. Inverse Lever Rule 12.5. Chromatography Scaleup 12.5.1. Background 12.5.2. Objectives 12.5.3. Procedure Report Appendix. Table of Units and Constants Index

Roger G. Harrison, University of Oklahoma, Paul Todd, Space Hardware Optimization Technology, Inc., Scott R. Rudge, FeRx, Inc., and Demetri P. Petrides, Intelligen, Inc.