DOWNSTREAM INDUSTRIAL BIOTECHNOLOGY -- CONTENTS -- PREFACE -- CONTRIBUTORS -- PART I INTRODUCTION -- 1 Bioprocess Design, Computer-Aided -- 1.1 INTRODUCTION -- 1.2 BENEFITS FROM THE USE OF COMPUTER AIDS -- 1.3 COMMERCIALLY AVAILABLE TOOLS -- 1.4 MONOCLONAL ANTIBODY EXAMPLE -- 1.5 DESIGN AND OPERATION OF MULTIPRODUCT FACILITIES -- 1.6 SUMMARY AND CONCLUSIONS -- REFERENCES -- PART II DOWNSTREAM RECOVERY OF CELLS AND PROTEIN CAPTURE -- 2 Cell Separation, Centrifugation -- 2.1 INTRODUCTION -- 2.2 CENTRIFUGAL SEPARATION -- 2.3 TYPES OF CENTRIFUGAL SEPARATORS -- 2.4 FLUID AND PARTICLE DYNAMICS -- 2.5 THE THEORETICAL SIZE OF A CENTRIFUGAL SEPARATOR -- 2.6 SELECTION OF TYPE AND SIZE OF CENTRIFUGE -- 2.7 DESCRIPTION OF SOME APPLICATIONS -- 2.8 INSTALLATION AND OPERATION -- 2.9 CENTRIFUGATION VERSUS MICROFILTRATION -- NOMENCLATURE -- REFERENCES -- 3 Cell Disruption, Micromechanical Properties -- 3.1 INTRODUCTION -- 3.2 MICROORGANISMS: COMPOSITION AND MORPHOLOGY -- 3.3 MICROMECHANICAL PROPERTIES OF MICROORGANISMS -- 3.4 CELL DISRUPTION -- 3.5 COMPARISON OF DIFFERENT CELL DISRUPTION DEVICES -- 3.6 CORRELATION OF MICROMECHANICAL RESULTS WITH CELL DISRUPTION RESULTS -- 3.7 GREEK LETTERS -- NOMENCLATURE -- REFERENCES -- 4 Cell Separation, Yeast Flocculation -- 4.1 INTRODUCTION -- 4.2 MICROBIAL AGGREGATION AND YEAST FLOCCULATION: SCOPE AND DEFINITIONS -- 4.3 GENETICS OF YEAST FLOCCULATION -- 4.4 MOLECULAR MECHANISM OF YEAST FLOCCULATION -- 4.5 INDUCTIVELY VERSUS CONSTITUTIVELY FLOCCULENT STRAINS -- 4.6 ENVIRONMENTAL FACTORS AFFECTING YEAST FLOCCULATION -- 4.7 YEAST FLOCCULATION AND BIOTECHNOLOGICAL PROCESSES -- REFERENCES -- 5 Cell Wall Disruption and Lysis -- 5.1 INTRODUCTION -- 5.2 THE CELL WALL -- 5.3 ASSESSMENT OF THE DISRUPTION YIELD -- 5.4 METHODS OF CELL WALL DISRUPTION -- 5.5 EFFECTS OF CELL DISRUPTION ON DOWNSTREAM OPERATIONS.

5.6 PROCESS INTENSIFICATION BY INTEGRATION OF DISRUPTION WITH PROTEIN CAPTURE -- REFERENCES -- 6 Expanded Bed Chromatography, Surface Energetics of Biomass Deposition -- 6.1 INTRODUCTION -- 6.2 TECHNICAL CHALLENGES IN EBA -- 6.3 SURFACE THERMODYNAMICS OF BIOMASS DEPOSITION IN EBA -- 6.4 SURFACE ENERGETICS AND PROTEIN ADSORPTION -- 6.5 CONCLUSION -- 6.6 NOMENCLATURE -- REFERENCES -- 7 Filter Aids -- 7.1 INTRODUCTION -- 7.2 PROCESS CLARIFICATION -- 7.3 POROUS MEDIA IN DYNAMIC PROCESS FILTRATIONS -- 7.4 FUNDAMENTAL PRINCIPLES OF DIATOMITE FILTRATION -- 7.5 GRADE SELECTION AND OPTIMIZATION -- 7.6 SYSTEMATIC METHODS DEVELOPMENT APPROACH TO GRADE SELECTION -- 7.7 SUMMARY -- REFERENCES -- 8 Protein Adsorption, Expanded Bed -- 8.1 INTRODUCTION -- 8.2 THEORY -- 8.3 PRINCIPLES OF OPERATION -- 8.4 EQUIPMENT -- 8.5 APPLICATIONS -- REFERENCES -- PART III PROCESS DEVELOPMENT IN DOWNSTREAM PURIFICATION -- 9 Scaledown of Biopharmacuetical Purification Operations -- 9.1 INTRODUCTION -- 9.2 GENERAL CONSIDERATIONS -- 9.3 CENTRIFUGATION -- 9.4 HOMOGENIZATION -- 9.5 REFOLDING -- 9.6 PRECIPITATION -- 9.7 CHROMATOGRAPHY -- 9.8 MICROFILTRATION AND ULTRAFILTRATION/DIAFILTRATION (UF/DF) -- 9.9 VIRAL CLEARANCE VIA CHROMATOGRAPHY AND FILTRATION -- 9.10 VIRAL INACTIVATION -- 9.11 MEMBRANE ADSORBERS -- 9.12 SUMMARY -- LIST OF ABBREVIATIONS -- NOMENCLATURE -- REFERENCES -- 10 Adsorption in Simulated Moving Beds (SMB) -- 10.1 INTRODUCTION -- 10.2 FUNDAMENTALS OF CHROMATOGRAPHIC SEPARATIONS -- 10.3 DESIGN OF OPERATING CONDITIONS -- 10.4 APPLICATIONS -- 10.5 ADVANCES OF SMB TECHNOLOGY -- 10.6 CONCLUDING REMARKS -- 10.7 NOMENCLATURE -- REFERENCES -- 11 Adsorption of Proteins with Synthetic Materials -- 11.1 INTERFACES -- 11.2 PROTEINS AT INTERFACES -- REFERENCES -- 12 Affinity Fusions for Protein Purification -- 12.1 INTRODUCTION -- 12.2 SYSTEMS FOR RAPID PROTEIN CAPTURE.

12.3 STABILIZATION OF EXPRESSED PROTEINS -- 12.4 DETECTION OF PRODUCED PROTEINS -- 12.5 REMOVAL OF AFFINITY TAGS -- 12.6 UTILIZATION OF FUSION PROTEINS AS ANTIGENS -- 12.7 SUBUNIT IMMUNOGENS FOR VACCINE RESEARCH -- 12.8 CONCLUSIONS -- REFERENCES -- 13 Bioseparation, Magnetic Particle Adsorbents -- 13.1 INTRODUCTION -- 13.2 SELECTED SCALABLE SYNTHESIS PROCEDURES -- 13.3 MAGNETIC ADSORBENTS FOR LABORATORY SEPARATIONS -- 13.4 MAGNETIC SEPARATION TECHNIQUES -- 13.5 SUMMARY -- REFERENCES -- 14 High Throughput Technologies in Bioprocess Development -- 14.1 INTRODUCTION -- 14.2 HTT APPLIED TO UPSTREAM CELL CULTURE DEVELOPMENT -- 14.3 HTT APPLIED TO DOWNSTREAM PURIFICATION DEVELOPMENT -- 14.4 ANALYSIS NEEDS FOR HIGH THROUGHPUT FORMATS -- 14.5 EXPERIMENTAL DESIGN FOR HTT -- 14.6 CONCLUSION -- REFERENCES -- 15 Large-Scale Protein Purification, Self-Cleaving Aggregation Tags -- 15.1 INTRODUCTION -- 15.2 CONVENTIONAL AFFINITY-TAG TECHNOLOGY -- 15.3 SELF-CLEAVING IN PROTEINS -- 15.4 CONVENTIONAL SELF-CLEAVABLE AFFINITY TAGS -- 15.5 SELF-CLEAVING AGGREGATION TAGS -- 15.6 ADVANTAGES, ECONOMY AND FUTURE PROSPECTS OF SELF-CLEAVING AGGREGATION-TAG TECHNOLOGIES -- REFERENCES -- 16 Lipopolysaccharide, LPS removal, Depyrogenation -- 16.1 INTRODUCTION -- 16.2 ENDOTOXINS: CHEMICAL AND PHYSICAL PROPERTIES -- 16.3 MECHANISM OF ENDOTOXIN ACTION -- 16.4 TECHNIQUES APPLIED FOR ENDOTOXIN REMOVAL -- 16.5 ENDOTOXIN REMOVAL IN BIOTECHNOLOGY MANUFACTURING PROCESSES -- REFERENCES -- 17 Porous Media in Biotechnology -- 17.1 INTRODUCTION -- 17.2 GENERAL DEFINITIONS -- 17.3 CHARACTERISTICS OF POROUS MEDIA -- 17.4 TRANSPORT PHENOMENA IN POROUS SYSTEMS -- 17.5 POROUS MEDIA IN BIOPROCESSES -- 17.6 CONCLUSION -- REFERENCES -- 18 Protein Aggregation and Precipitation, Measurement and Control -- 18.1 INTRODUCTION.

18.2 COMBINING METHODS TO FOLLOW AGGREGATION AND PRECIPITATION AND DETERMINE THE STRUCTURE OF COMPLEXES -- 18.3 SPECTRAL METHODS FOR MEASURING SOLUBILITY AND PROTEIN ASSOCIATION -- 18.4 UNDERSTANDING PROTEIN-SOLVENT INTERACTIONS PROTEIN STABILITY IN A PRACTICAL SENSE -- 18.5 DETERMINING THE SURFACE CHARGE AND HYDROPHOBICITY OF A PROTEIN -- 18.6 EMPIRICAL MODELS FOR SALTING IN AND PRECIPITATION WITH VARIOUS AGENTS -- 18.7 MODELS FOR DETERMINING THE EFFECT OF CO-SOLVENTS ON PROTEIN FOLDING -- 18.8 COMPUTER DESIGN OF MORE SOLUBLE PROTEINS -- 18.9 AUTOMATIC HOMOLOGY MODELING -- 18.10 MODELING USING SELF-CORRECTING DISTANCE GEOMETRY WITH THE PROGRAMS CLUSTAL, MASIA, NOAH, DIAMOD, AND FANTOM, TO DEVELOP A 3D MODEL OF A PROTEIN -- 18.11 CONCLUSIONS -- REFERENCES -- PART IV EQUIPMENT DESIGN FOR DOWNSTREAM RECOVERY AND PROTEIN PURIFICATION -- 19 Cleaning and Sanitation in Downstream Processes -- 19.1 INTRODUCTION -- 19.2 DESIGNING AN EFFECTIVE CLEANING PROTOCOL FOR DOWNSTREAM BIOPROCESSES -- 19.3 CHROMATOGRAPHIC MEDIA -- 19.4 CROSS-FLOW FILTRATION (CFF) -- 19.5 EQUIPMENT -- 19.6 SANITIZATION AND STERILIZATION -- 19.7 CLEANING VALIDATION -- 19.8 CONCLUSIONS -- REFERENCES -- 20 Clean-in-place -- 20.1 INTRODUCTION -- 20.2 THE REQUIREMENT FOR CIP SYSTEMS -- 20.3 GENERAL OUTLINE OF A CIP REGIME -- 20.4 CIP CHEMICALS -- 20.5 CIP DESIGN AND CONSTRUCTION -- 20.6 CIP CONFIGURATION -- 20.7 AUTOMATION -- 20.8 VALIDATION AND VERIFICATION -- REFERENCES -- FURTHER READING -- 21 Large Scale Chromatography Columns, Modeling Flow Distribution -- 21.1 INTRODUCTION -- 21.2 CHALLENGES OF SCALING UP CHROMATOGRAPHY -- 21.3 ANALYSIS ON THE WALL EFFECT -- 21.4 MODEL THE COUPLING BETWEEN BED COMPRESSION AND FLOW -- 21.5 IMPACT OF HARDWARE DESIGN ON THE FLOW IN LARGE COLUMNS -- 21.6 MODELING THE TRANSPORT OF ELUTION AND HETP ANALYSIS -- 21.7 SUMMARY -- LIST OF ABBREVIATIONS.

NOMENCLATURE -- REFERENCES -- 22 Pumps, Industrial -- 22.1 INTRODUCTION -- 22.2 THEORY -- 22.3 CENTRIFUGAL PUMPS -- 22.4 POSITIVE DISPLACEMENT PUMPS -- 22.5 DRIVERS -- 22.6 SPECIAL CONSIDERATIONS FOR BIOPROCESSING PUMPS -- 22.7 TROUBLESHOOTING -- ADDITIONAL READING -- PART V DOWNSTREAM cGMP OPERATIONS -- 23 Affinity Chromatography of Plasma Proteins -- 23.1 INTRODUCTION -- 23.2 LIGANDS AND SUPPORTS FOR AFFINITY PURIFICATION -- 23.3 APPLICATION OF AFFINITY CHROMATOGRAPHY IN PLASMA PROTEIN PROCESSES -- 23.4 QUALITY CONTROL OF AFFINITY-PURIFIED PROTEINS -- 23.5 CONCLUSIONS -- REFERENCES -- 24 Antibody Purification, Monoclonal and Polyclonal -- 24.1 INTRODUCTION -- 24.2 APPROACH TO DOWNSTREAM PROCESSING -- 24.3 AFFINITY CHROMATOGRAPHY -- 24.4 ION-EXCHANGE CHROMATOGRAPHY -- 24.5 HYDROPHOBIC INTERACTION CHROMATOGRAPHY -- 24.6 CERAMIC HYDROXYAPATITE CHROMATOGRAPHY -- 24.7 MIXED MODE CHROMATOGRAPHY -- 24.8 PURIFICATION OF IgM -- 24.9 PLATFORM PROCESSES -- 24.10 CONCLUSION -- REFERENCES -- 25 Chromatographic Purification of Virus Particles -- 25.1 INTRODUCTION -- 25.2 CHROMATOGRAPHIC SEPARATION METHODS -- 25.3 ADSORPTION CHROMATOGRAPHY -- 25.4 ION EXCHANGE CHROMATOGRAPHY -- 25.5 HYDROPHOBIC INTERACTIONS CHROMATOGRAPHY -- 25.6 MULTIMODAL METHODS -- 25.7 OTHER MULTIMODAL METHODS -- 25.8 BIOSPECIFIC AFFINITY CHROMATOGRAPHY -- 25.9 PROCESS DEVELOPMENT -- 25.10 SAMPLE DEFINITION -- 25.11 SAMPLE PREPARATION -- 25.12 INITIAL SCREENING -- 25.13 BIOSPECIFIC AFFINITY -- 25.14 INTERPRETATION OF INITIAL RESULTS -- 25.15 CONCLUDING REMARKS -- 25.16 RECOMMENDED READING -- REFERENCES -- 26 Chromatography, Hydrophobic Interaction -- 26.1 INTRODUCTION -- 26.2 HYDROPHOBIC INTERACTION -- 26.3 HYDROPHOBIC INTERACTION CHROMATOGRAPHY -- 26.4 CLASSIFYING OF MEDIA AND MODELLING OF CHROMATOGRAPHIC RESULTS -- 26.5 THE CHROMATOGRAPHY CONDITIONS.

26.6 REGENERATION AND CLEANING-IN-PLACE.