Downstream Processing

  1. Stage 4: Purification by Chromatography
    1. Fundamentals of Chromatography
      1. Basic Principles
        1. Stationary Phase Properties
          1. Mobile Phase Selection
            1. Retention Mechanisms
            2. Performance Parameters
              1. Resolution
                1. Selectivity
                  1. Efficiency
                    1. Capacity
                    2. Adsorption Isotherms
                      1. Langmuir Isotherm
                        1. Freundlich Isotherm
                          1. Linear Isotherm
                          2. Mass Transfer Effects
                            1. Band Broadening
                              1. Van Deemter Equation
                            2. Ion-Exchange Chromatography
                              1. Principles of Ion Exchange
                                1. Electrostatic Interactions
                                  1. pH Effects on Binding
                                    1. Ionic Strength Effects
                                    2. Anion Exchange
                                      1. Strong Anion Exchangers
                                        1. Weak Anion Exchangers
                                          1. Common Resins and Ligands
                                          2. Cation Exchange
                                            1. Strong Cation Exchangers
                                              1. Weak Cation Exchangers
                                                1. Common Resins and Ligands
                                                2. Process Development
                                                  1. pH Scouting
                                                    1. Salt Gradient Optimization
                                                      1. Loading Conditions
                                                      2. Elution Strategies
                                                        1. Step Elution
                                                          1. Gradient Elution
                                                            1. pH Elution
                                                          2. Affinity Chromatography
                                                            1. Principles of Molecular Recognition
                                                              1. Specific Binding Interactions
                                                                1. Ligand Selection
                                                                  1. Binding Kinetics
                                                                  2. Protein A Chromatography
                                                                    1. Mechanism of Antibody Binding
                                                                      1. Process Conditions
                                                                        1. Elution Methods
                                                                        2. Protein G Chromatography
                                                                          1. Comparison with Protein A
                                                                          2. Immobilized Metal Affinity Chromatography
                                                                            1. Metal Selection
                                                                              1. His-tag Purification
                                                                                1. Process Optimization
                                                                                2. Other Affinity Systems
                                                                                  1. Heparin Affinity
                                                                                    1. Lectin Affinity
                                                                                      1. Dye Ligand Affinity
                                                                                      2. Elution Strategies
                                                                                        1. Competitive Elution
                                                                                          1. pH Shift Elution
                                                                                            1. Chaotropic Elution
                                                                                          2. Hydrophobic Interaction Chromatography
                                                                                            1. Principles of Hydrophobic Interactions
                                                                                              1. Salt Effects and Lyotropic Series
                                                                                                1. Resin Selection
                                                                                                  1. Process Development
                                                                                                    1. Salt Screening
                                                                                                      1. Gradient Optimization
                                                                                                        1. Temperature Effects
                                                                                                        2. Applications and Limitations
                                                                                                        3. Size-Exclusion Chromatography
                                                                                                          1. Principles of Size Separation
                                                                                                            1. Column Selection and Packing
                                                                                                              1. Applications in Polishing
                                                                                                                1. Aggregate and Fragment Removal
                                                                                                                  1. Buffer Selection
                                                                                                                    1. Flow Rate Optimization
                                                                                                                    2. Reversed-Phase Chromatography
                                                                                                                      1. Principles of Hydrophobic Interactions
                                                                                                                        1. Stationary Phase Selection
                                                                                                                          1. Mobile Phase Considerations
                                                                                                                            1. Applications for Small Molecules
                                                                                                                              1. Applications for Peptides
                                                                                                                                1. Gradient Development
                                                                                                                                2. Mixed-Mode Chromatography
                                                                                                                                  1. Principles of Multiple Interactions
                                                                                                                                    1. Resin Types and Properties
                                                                                                                                      1. Process Development
                                                                                                                                        1. Applications and Advantages
                                                                                                                                        2. Chromatography System Design
                                                                                                                                          1. Column Design and Packing
                                                                                                                                            1. Packing Techniques
                                                                                                                                              1. Column Qualification
                                                                                                                                                1. Performance Testing
                                                                                                                                                2. System Components
                                                                                                                                                  1. Pumps and Flow Control
                                                                                                                                                    1. Detection Systems
                                                                                                                                                      1. Fraction Collection
                                                                                                                                                      2. Process Control
                                                                                                                                                        1. Gradient Formation
                                                                                                                                                          1. Flow Rate Control
                                                                                                                                                            1. Pressure Monitoring
                                                                                                                                                            2. Operating Modes
                                                                                                                                                              1. Bind-and-Elute Mode
                                                                                                                                                                1. Flow-Through Mode
                                                                                                                                                                  1. Displacement Chromatography
                                                                                                                                                                2. Cleaning and Maintenance
                                                                                                                                                                  1. Cleaning-in-Place Procedures
                                                                                                                                                                    1. Sanitization-in-Place Procedures
                                                                                                                                                                      1. Cleaning Agent Selection
                                                                                                                                                                        1. Validation of Cleaning
                                                                                                                                                                          1. Resin Lifetime and Storage

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                                                                                                                                                                        4. Stage 3: Concentration and Initial Purification

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                                                                                                                                                                        6. Stage 5: Final Purification and Polishing