Preface xix 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 Stability of Small Biomolecules 1.3.4 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 1.5.1 Structure of Nucleic Acids 1.5.2 Functions and Commercial Uses 1.5.3 Stability of Nucleic Acids 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 1.10 Summary Nomenclature Problems References 2. Analytical Methods and Bench Scale Preparative Bioseparations 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 Ultraviolet 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 (AES) 2.5.12 Dry Weight 2.5.13 Flow Cytometry 2.

6 Microbiology Assays 2.6.1 Sterility 2.6.2 Bioburden 2.6.3 Endotoxin 2.6.

4 Virus, Mycoplasma, and Phage 2.7 Bench Scale Preparative Separations 2.7.1 Preparative Electrophoresis 2.7.2 Magnetic Bioseparations 2.7.3 Cell Purification by Flow Cytometry 2.

8 Summary Nomenclature Problems References 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 Counterion Charge Number 3.4.3 The Schulze-Hardy Rule 3.

4.4 Flocculation Rate 3.4.5 Polymeric Flocculants 3.5 Summary Nomenclature Problems References 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 Example 4.

3. Comparison of Mass Transfer Coefficient Calculated by Boundary Layer Theory Versus by Shear-Induced Diffusion Theory 4.3 Filter Media and Equipment 4.3.1 Conventional Filtration 4.3.2 Crossflow Filtration 4.4 Membrane Fouling 4.

5 Scale-up and Design of Filtration Systems 4.5.1 Conventional Filtration Example 4.4. Rotary Vacuum Filtration Example 4.5. Washing of a Rotary Vacuum Filter Cake 4.5.

2 Crossflow Filtration Example 4.6. Diafiltration Mode in Crossflow Filtration 4.6 Summary Nomenclature Problems References 5. Sedimentation 5.1 Instructional Objectives 5.2 Sedimentation Principles 5.2.

1 Equation of Motion 5.2.2 Sensitivities 5.3 Methods for Analysis of Sedimentation 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. Scale-up 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 Péclet Analysis 5.7.4 Inclined Sedimentation 5.7.5 Field-Flow Fractionation 5.8 Centrifugal Elutriation 5.9 Summary Nomenclature Problems References 6.

Extraction 6.1 Instructional Objectives 6.2 Extraction Principles 6.2.1 Phase Separation and Partitioning Equilibria Example 6.1 Process for Large-Scale Isolation of ?-Galactosidae from E. coli in an Aqueous Two-Phase Sytstem 6.2.

2 Countercurrent Stage Calculations Example 6.2. Separation of a Bioproduct and an Impurity by Countercurrent Extraction Example 6.3. Effect of Solvent Rate in Countercurrent Staged Extraction of an Antibiotic 6.3 Scale-up and Design of Extractors 6.3.1 Reciprocating-Plate Extraction Columns Example 6.

4. Scale-up of a Reciprocating-Plate Extraction Column 6.3.2 Centrifugal Extractors Example 6.5. Increase in Feed Rate to a Podbielniak Centrifugal Extractor 6.4 Summary Nomenclature Problems References 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 Data 7.3.

2 Agitated-Bed Adsorption 7.4 Chromatography Column Dynamics 7.4.1 Plate Models 7.4.2 Moment Analysis Example 7.2 Calculation of the HETP Using the Method of Moments 7.4.

3 Chromatography Column Mass Balance with Negligible Dispersion Example 7.3. Chromatographic Separation of Two Solutes Example 7.4. Calculation of the Shock Wave Velocity for a Nonlinear Isotherm Example 7.5. Calculation of the Elution Profile 7.4.

4 Dispersion Effects in Chromatography 7.4.5 Computer Simulation of Chromatography Considering Axial Dispersion, Fluid-Phase Mass Transfer, Intraparticle Diffusion, and Nonlinear Equilibrium Example 7.6 Computer Simulation of a Chromatography Process 7.4.6 Gradients and Modifiers Example 7.7. Equilibrium for a Protein Anion in the Presence of Chloride Ion 7.

5 Membrane Chromatography Example 7.8. Comparison of Time for Diffusion Mass Transfer in Conventional Chromatography and Membrane Chromatography 7.6 Simulated Moving Bed Chromatography 7.7 Adsorbent Types 7.7.1 Silica-Based Resins 7.7.

2 Polymer-Based Resins 7.7.3 Ion Exchange Chromatography and Adsorption 7.7.4 Reversed-Phase Chromatography 7.7.5 Hydrophobic Interaction Chromatography 7.7.

6 Affinity Chromatography 7.7.7 Immobilized Metal Affinity Chromatography (IMAC) 7.7.8 Size Exclusion Chromatography 7.8 Particle Size and Pressure Drop in Fixed Beds 7.9 Equipment 7.9.

1 Columns 7.9.2 Chromatography Column Packing Procedures 7.9.3 Detectors 7.9.4 Chromatography System Fluidics 7.10 Scale-up 7.

10.1 Adsorption Example 7.9. Scale-up of the Fixed-Bed Adsorption of a Pharmaceutical Product 7.10.2 Chromatography Example 7.10. Scale-up of a Protein Chromatography Example 7.

11. Scale-up of Protein Chromatography Using Standard Column Sizes Example 7.12. Scale-up of Elution Buffer Volumes in Protein Chromatography Example 7.13. Consideration of Pressure Drop in Column Scaling 7.11 Summary Nomenclature Problems References 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 Precipitate 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.