Table of Contents

CHAPTER 1: INTRODUCTION
     1.1.The Complexity of Biological Materials
          1.1.1.Starting Sample Complexity
          1.1.2.General Bioseparation Schema
     1.2.Translating Structure to Exploitable Properties
     1.3.The Basis and Relative Cost of Bioseparation Processes
          1.3.1.Cost of Continuous Processes
          1.3.2.Cost of Batch Processes
CHAPTER 2: BIOMOLECULES
     2.1.The Major Biochemical Components
     2.2.Water
          2.2.1.Water as a Solvent
          2.2.2.Water as a Source of Ions
          2.2.3.Oxidation-Reduction Reactions with Water
     2.3. Small Molecule Pools
          2.3.1.Fatty Acids, and Membrane-Soluble Materials
          2.3.2.Primary and Secondary Metabolites
          2.3.3.Amino Acids
          2.3.4.Nucleosides and Nucleotides
          2.3.5.Sugars
     2.4.Macromolecules
          2.4.1.Lipids
          2.4.2.Polysaccharides
          2.4.3.Lipopolysaccharides and Peptidoglycan
          2.4.4.Proteins
          2.4.5.Nucleic Acids
CHAPTER 3: SOLUBILITY
     3.1.Solvent-Solute Interactions
          3.1.1.London Dispersion Force (Van der Waals Interactions)
          3.1.2.Dipole Interactions
          3.1.3.Hydrogen Bonding
          3.1.4.Ions
          3.1.5.Size Matters
     3.2.Solubility Parameters
          3.2.1.Hansch Lipophilic Parameter
          3.2.2.Hildebrand Parameter
          3.2.3.Hansen Parameters
          3.2.4.Hansen Parameter Estimation by Group Contribution Method
     3.3.Applications of Solubility Parameters
          3.3.1.The Solubility of Cellular Components
          3.3.2.Phase Equilibria
          3.3.3.Macromolecular Biomolecule Solubility
          3.3.4.Mobile Phase Optimization in Chromatography
CHAPTER 4: MASS AND SIZE
     4.1.Atomic Composition
          4.1.1.Elemental Mass
          4.1.2.Nuclear Binding Energy
          4.1.3.Molecules
          4.1.4.Isotopic Distribution
          4.1.5.Mass Changes on Ionization
          4.1.6.Radicals and Radical Ions
     4.2.Molecular size
          4.2.1.Molar Volume
          4.2.2.Volume Change on Mixing
          4.2.3.Diffusivity
          4.3.4.Rotational Diffusivity
          4.3.5.Radii of Gyration and Hydration
          4.3.6.MW Distribution
     4.3.Comparative Sizes
CHAPTER 5: IONIZATION
     5.1.Ionization Equilibria
          5.1.1.Simple Acids and Bases
          5.1.2.Multiple-Ionizable Groups
          5.1.3.Total and Net Charge
          5.1.4.Isoelectric Point
     5.2.Electroneutrality Constraint
     5.3.Buffer Capacity
CHAPTER 6: PRECIPITATION AND CRYSTALIZATION
     6.1.Precipitation and Crystallization Mechanisms
          6.1.1.Thermal Cycling
          6.1.2.Ionic Solubility
          6.1.3.Miscible Solvents
          6.1.4.Precipitation with Polymers
          6.1.5.Solvent Volume Reduction
     6.2.Particle Growth
          6.2.1.Nucleation
          6.2.2.Particle Growth
          6.2.3.Particle Diameter Growth
          6.2.4.Particle Size Distribution
     6.3.Protein Precipitation and Crystallization
          6.3.1.Protein Precipitation
          6.3.2.Protein Crystallization
CHAPTER 7: EXTRACTION AND LEACHING
     7.1.Phase Equilibrium
          7.1.1.The Thermodynamics of Phase Equilibrium
          7.1.2.Mass Transport Kinetics
          7.1.3.Extraction Efficiency
     7.2.Extractions
          7.2.1. Batch Extraction
          7.2.2. Continuous Extraction
     7.3.Leaching
          7.3.1.Mass Transfer from the Solid Particle
          7.3.2.Fixed Bed Leaching Process
          7.3.3.Pressure Drop and Particle Stability During Fixed-Bed Leaching
CHAPTER 8: CHROMATOGRAPHY
     8.1.Conceptual Chromatography Model
          8.1.1.Mobile Phase Model
          8.1.2.Boundary and Initial Conditions
     8.2.A More General Model
          8.2.1.Mobile Phase Through a Bead-Packed Column
          8.2.2.Model of the Solid Surface (Bead)
          8.2.3.Dimensional Analysis of the Model
     8.3.Estimation and Use of Dimensionless Parameters
          8.3.1.Dimensional Parameters
          8.3.2.Column Pressure Drop Constraint
     8.4.Langmuir Isotherm
          8.4.1.Adsorption versus Absorption
          8.4.2.An Illustrative Thought Experiment
          8.4.3.Langmuir Isotherm
          8.4.4.Measuring the Binding Affinity and Number of Binding Sites
          8.4.5.Langmuir Isotherm Based on the Total Analyte Concentration
          8.4.6.Multicomponent Langmuir Isotherm
     8.5.Chromatography Process Scale-Up
          8.5.1.Resolving Power
          8.5.2.Scale-Up
CHAPTER 9: CHROMATOGRAPHIC MODES
     9.1.Reverse Phase and Hydrophobic Interaction
          9.1.1.Mobile Phase
          9.1.2.Solid Phase
          9.1.3.Mobile Phase Modification
     9.2.Ion Exchange Chromatography
          9.2.1.Types of Ion Exchange Resins
          9.2.2.Ion Exchange
          9.2.3.pH Isotherm
          9.2.4.Ion Displacement Isotherm
          9.2.5.Polyionic Analytes
     9.3.Size Exclusion Chromatography
          9.3.1.Measuring the Apparent Porosity
          9.3.2.Effective Diffusivity Inside Pores
          9.3.3.SEC Applications
     9.4.Affinity Chromatography
     9.5.Displacement Chromatography
          9.5.1.Displacement Mechanism Model
          9.5.2.Displacing Agents
CHAPTER 10: ELECTROPHORESIS
     10.1.The Electrophoretic Process
          10.1.1.Process Description
          10.1.2.Electrochemical Thermodynamics
     10.2.Rigorous Model for Electrophoresis
          10.2.1.Material Balance Equations
          10.2.2.Phenomenological Equations
          10.2.3.Model for Total Chemical Component
          10.2.4.Current
          10.2.5.Local Electroneutrality Constraint
          10.2.6. Numerical Solutions
     10.3.Joule Heating
     10.4.A Practical Model
     10.5. Electroosmotic Flow
          10.5.1.Charged Surface of the Electrolyte Channel
          10.5.2.Electroosmotic Flow (EOF)
          10.5.3.Effect of EOF on Separations
          10.5.4.Apparent Mobility
          10.5.5.Design for Separation
CHAPTER 11: ELECTROPHORETIC SEPARATION MODES
     11.1.Isotachophoresis
     11.2.Isoelectric focusing (IEF)
          11.2.1pH Gradient
          11.2.2.Maintaining the pH Gradient
          11.2.3.Isoelectric Precipitation
          11.2.4.Continuous Isoelectric Focusing
     11.3Zone Electrophoresis
          11.3.1.Electrophoretic Mobility and Molecular Ion Size
          11.3.2.Separation by Electrophoretic Mobility
          11.3.3.Joule Heating Limitation in Zone Electrophoresis
          11.3.4.Electroosmotic Flow
     11.4.Gel Electrophoresis
          11.4.1.Sieving Potential
          11.4.2. Gel Electrophoretic Applications
          11.4.3.Process-Scale Gel Electrophoresis
     11.5.Electrodialysis246
          11.5.1.Electrophoretic Rate Enhancement
          11.5.2.Precipitation at the Membrane
          11.5.3.Scale-up Considerations
          11.5.4.Continuous Electrodialysis
     11.6.Electrochromatography
CHAPTER 12: PARTICULATE-LIQUID SEPARATIONS
     12.1.The Nature of Biological Particles
     12.2.Filtration
          12.2.1.Filter Types
          12.2.2.Filtration Modes
          12.2.3.Pressure Drop through a Filter
     12.3.Centrifugation
          12.3.1.Stokes’ Law
          12.3.2.Correction for Particle Interactions
          12.3.3.Speed and Time Requirements of a Centrifuge
          12.3.4.Batch and Continuous Centrifugation
     12.4.Basket Centrifuge or Centrifugal filtration
          12.4.1.Liquid Velocity and Pressure Drop
          12.4.2.Force on the Solid Particles
          12.3.3.Empty Bed Case
          12.3.4.Centrifuge Design Constraints
APPENDIX A: TABLE OF HANSEN PARAMETER VALUES
APPENDIX B: FRACTIONAL PARAMETER VALUES FOR GROUP CONTRIBUTION ESTIMATION OF HANSEN SOLUBILITY PARAMETERS
APPENDIX C: SET OF TEST SOLVENTS USEFUL FOR DEFINING THE HANSEN SOLUBILITY ENVELOPE
APPENDIX D: ISOTPIC MASSES AND NATURAL ABUNDANCES