Downstream Industrial Biotechnology E-Book

• Herausgeber: John Wiley & Sons
• Kategorie: Fachliteratur
• Sprache: Englisch

Seitenzahl: 2441

Veröffentlichungsjahr: 2013

Beschreibung

DOWNSTREAM INDUSTRIAL BIOTECHNOLOGY An affordable, easily accessible desk reference on biomanufacturing, focused on downstream recovery and purification Advances in the fundamental knowledge surrounding biotechnology, novel materials, and advanced engineering approaches continue to be translated into bioprocesses that bring new products to market at a significantly faster pace than most other industries. Industrial scale biotechnology and new manufacturing methods are revolutionizing medicine, environmental monitoring and remediation, consumer products, food production, agriculture, and forestry, and continue to be a major area of research. The downstream stage in industrial biotechnology refers to recovery, isolation, and purification of the microbial products from cell debris, processing medium and contaminating biomolecules from the upstream process into a finished product such as biopharmaceuticals and vaccines. Downstream process design has the greatest impact on overall biomanufacturing cost because not only does the biochemistry of different products ( e.g., peptides, proteins, hormones, antibiotics, and complex antigens) dictate different methods for the isolation and purification of these products, but contaminating byproducts can also reduce overall process yield, and may have serious consequences on clinical safety and efficacy. Therefore downstream separation scientists and engineers are continually seeking to eliminate, or combine, unit operations to minimize the number of process steps in order to maximize product recovery at a specified concentration and purity. Based on Wiley's Encyclopedia of Industrial Biotechnology: Bioprocess, Bioseparation, and Cell Technology, this volume features fifty articles that provide information on down- stream recovery of cells and protein capture; process development and facility design; equipment; PAT in downstream processes; downstream cGMP operations; and regulatory compliance. It covers: * Cell wall disruption and lysis * Cell recovery by centrifugation and filtration * Large-scale protein chromatography * Scale down of biopharmaceutical purification operations * Lipopolysaccharide removal * Porous media in biotechnology * Equipment used in industrial protein purification * Affinity chromatography * Antibody purification, monoclonal and polyclonal * Protein aggregation, precipitation and crystallization * Freeze-drying of biopharmaceuticals * Biopharmaceutical facility design and validation * Pharmaceutical bioburden testing * Regulatory requirements Ideal for graduate and advanced undergraduate courses on biomanufacturing, biochemical engineering, biopharmaceutical facility design, biochemistry, industrial microbiology, gene expression technology, and cell culture technology, Downstream Industrial Biotechnology is also a highly recommended resource for industry professionals and libraries.

Leseprobe

Table of Contents

Title Page

Copyright

Preface

Contributors

Part I: Introduction

Introduction

Chapter 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

Chapter 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

2.10 Nomenclature

References

Chapter 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 Nomenclature

3.8 Greek Letters

References

Chapter 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

4.8 Conclusions

References

Chapter 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

Chapter 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

Further Reading

Chapter 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

Chapter 8: Protein Adsorption, Expanded Bed

8.1 Introduction

8.2 Theory

8.3 Principles of Operation

8.4 Equipment

8.5 Applications

References

Further Reading

Part III: Process Development in Downstream Purification

Chapter 9: Scaledown Of Biopharmaceutical 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

9.13 List Of Abbreviations

9.14 Nomenclature

References

Chapter 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

Further reading

Chapter 11: Adsorption of Proteins with Synthetic Materials

11.1 Interfaces

11.2 Proteins at Interfaces

References

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Summary

List of Abbreviations

Nomenclature

References

Chapter 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

Chapter 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

Chapter 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

Chapter 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

Chapter 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

26.7 Optimization Procedures

26.8 Applications

References

Chapter 27: Chromatography, Radial Flow

27.1 Introduction

27.2 Radial Flow Column Configurations

27.3 Packing Procedures for RFC Columns

27.4 Pressure Drops of RFC Columns

27.5 Comparison of Radial- and Axial Flow Columns

27.6 Pros and Cons of RFC Columns

27.7 Application Examples

27.8 Mathematical Modeling of Radial Flow Chromatography

27.9 Scale-Up of RFC Columns

27.10 Conclusions

Nomenclature

Greek Letters

References

Chapter 28: Drying, Biological Materials

28.1 Introduction

28.2 Drying of Biotechnological Products

28.3 Effect of Drying on Biotechnological Product Quality

28.4 Basic Principle of Drying

28.5 Commonly Used Dryers

28.6 Some Emerging Drying Technologies

28.7 Closing Remarks

References

Chapter 29: Freeze-Drying, Pharmaceuticals

29.1 Introduction

29.2 Pharmaceutical Freeze-Drying: Fundamentals

29.3 Challenges and New Advancements in Freeze-Drying

References

Chapter 30: Freezing, Biopharmaceutical

30.1 Introduction

30.2 Freezing of Solutions

30.3 Thawing of Solutions

30.4 Freeze-Thaw Scale Up

30.5 Conclusion

References

Chapter 31: Membrane Chromatography

31.1 Introduction

31.2 The Basic Concept

31.3 Limiting Factors in Membrane Adsorption Processes

31.4 Optimizing The Performance of Adsorptive Membranes

31.5 POSITIONING OF MEMBRANE ADSORPTION IN A PURIFICATION TRAIN

31.6 Applications

31.7 Concluding Remarks

Nomenclature

References

Chapter 32: Membrane Separations

32.1 Membrane Separations

32.2 Introduction

32.3 Three major applications of membrane separations

32.4 Classification of Membranes and Membrane Processes

32.5 Membrane Chemistry, Structure and Function

32.6 Methods of Membrane Manufacture

32.7 How Membrane Processes are Operated

32.8 Conclusion

References

Chapter 33: Plasmid Purification

33.1 Introduction

33.2 Therapeutic Plasmids

33.3 Cell Lysis

33.4 Chromatographic Methods

33.5 Nonchromatographic Methods

33.6 Industrial Processes

33.7 Conclusions and Perspectives

References

Chapter 34: Protein Chromatography, Manufacturing Scale

34.1 Introduction

34.2 Scale-Up of Chromatography

34.3 Impact of Chromatographic Parameters at Manufacturing Scale

34.4 Practical Issues Regarding Chromatography at Manufacturing Scale

34.5 Basic Chromatography Setup

References

Chapter 35: Protein Crystallization, Kinetics

35.1 Protein Molecules in Solution

35.2 Homogeneous Nucleation

35.3 Heterogeneous Nucleation

35.4 Nonclassical Approach to Nucleation

35.5 Crystal Growth

35.6 Crystallization in Forced Solution Flow Regime

References

Chapter 36: Protein Purification, Aqueous Liquid Extraction

36.1 Introduction

36.2 Biochemical Fundamentals

36.3 Alternative Two-Phase Systems

36.4 Applications

36.5 Conclusion

References

Chapter 37: Protein Ultrafiltration

37.1 Introduction

37.2 Theoretical Principles

37.3 Membrane Materials, Characterization, and Fouling

37.4 Modules and Devices

37.5 Equipment

37.6 Process Configurations

37.7 Process Design—Ultrafiltration

37.8 High Performance Tangential Flow Filtration

37.9 Scale-Up

Nomenclature

References

Chapter 38: Virus Retentive Filters

38.1 Process Virology Overview

38.2 Principles of Operation

38.3 Physical Attributes

38.4 Process Considerations

38.5 Filtration Modes

38.6 Filter Classes

38.7 Integrity Testing

38.8 Performance

38.9 Validation (Virus Clearance Evaluation) Studies

38.10 Future Trends

38.11 Concluding Comments

List of Abbreviations

References

Part VI: Biopharmaceutical Facility Validation

Chapter 39: Biopharmaceutical Facility Design and Validation

39.1 Introduction

39.2 Designing for Compliance

39.3 Risk Management

39.4 Qualification/Verification

39.5 Process Validation

List of Abbreviations

References

Chapter 40: Closed Systems in Bioprocessing

40.1 Introduction

40.2 Definition of Closed Systems

40.3 Closed System Design

40.4 Impact on Facility Design

40.5 Impact on Operations

40.6 Summary

References

Chapter 41: Facility Design for Single Use (SU) Downstream Materials

41.1 Introduction

41.2 Facility Design

41.3 Cell Culture

41.4 Purification

41.5 Applications in A Fill and Finish Plant

41.6 Media Buffer

41.7 Costs

41.8 Conclusions

References

Chapter 42: Cgmps For Production Rooms

42.1 General Points for Facility Design

42.2 Design of HVAC System Parameters

References

Additional Reading

Chapter 43: Heating, Ventilation, and Air Conditioning

43.1 Introduction

43.2 HVAC Design Process

43.3 Regulation and Code Consideration

43.4 Temperature, Humidity, and Airflow

43.5 Pressurization

43.6 Air Handling Systems

43.7 Dehumidifiers

43.8 Humidifiers

43.9 Air Handling Unit Application

43.10 Return and Exhaust Fan Selection and Locations

43.11 HEPA Filters

43.12 Terminal Air Control Devices

43.13 Air Terminal Outlets

43.14 Duct Materials, Pressure, and Cleanability

43.15 System Operating Procedures

43.16 Emergency Electrical Power

43.17 Building Control and Automation Systems

43.18 Testing, Balancing, and Cleaning

43.19 Validation

43.20 Summary

References

Chapter 44: Sterilization-in-Place (SIP)

44.1 Introduction

44.2 Applications

44.3 Steam-in-Place Technology

44.4 Alternative SIP Technologies

References

Part VII: FDA cGMP Regulatory Compliance

Chapter 45: Pharmaceutical Bioburden Testing

45.1 Introduction

45.2 Bioburden Considerations

45.3 Standard Industry Methodologies

45.4 New and Rapid Technologies

45.5 Conclusion

References

Further Reading

Chapter 46: Chromatography, Industrial Scale Validation

46.1 Introduction

46.2 Systems Requirements

46.3 CFR Part 11 Software Evaluation Checklist for Closed Systems which do not use Biometrics

46.4 Gmp Requirements

46.5 Conclusion

Chapter 47: GMPs and GLSPs

47.1 Introduction

47.2 Facility Regulatory Guidelines

47.3 Major Focuses of Facility Performance Validation

47.4 Investigations Of Nonconformances And Deviations

Chapter 48: Quality by Design (QBD)

48.1 Biopharmaceutical Development

48.2 Key Steps for Implementing QbD

48.3 Quality Risk Management for Biopharmaceuticals

48.4 Design Space(S) for Biopharmaceuticals

48.5 PAT in Biopharmaceutical Manufacture

48.6 Hypothetical QbD Case Study for a Biopharmaceutical: Anion Exchange Chromatography

48.7 FDA's QbD Pilot Program for Biopharmaceuticals

48.8 Summary

48.9 List of Abbreviations

Chapter 49: Regulatory Requirements, European Community

49.1 The European Union

49.2 The European Medicines Agency (EMEA)

49.3 New Drug Approval Routes Coordinated by the Emea

49.4 The Emea in the Context of Drug Development and Manufacture

49.5 Concluding Remarks

References

Index

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Library of Congress Cataloging-in-Publication Data:

Encyclopedia of industrial biotechnology. Selections.

Downstream industrial biotechnology : recovery and purification / edited by Michael C. Flickinger.

pages cm

Includes bibliographical references and index.

ISBN 978-1-118-13124-4 (hardback)

1. Biotechnology-Encyclopedias. I. Flickinger, Michael C., editor of compilation. II. Title.

TP248.16.E533 2013

660.6--dc23

2012030526

Preface

Downstream Industrial Biotechnology is a compilation of essential in depth articles, organized topically and listed in alphabetical format, for biopharmaceutical, bioprocess and biologics process scientists, engineers and regulatory professionals from the comprehensive seven volumes of the Encyclopedia of Industrial Biotechnology. Process development for the manufacture of complex biomolecules involves solving many scientific, compliance and technical problems quickly in order to support pilot, preclinical and clinical development, technology transfer and manufacturing start-up. Every organization develops new processes from accumulated process knowledge. Accumulated process knowledge has a very significant impact on accelerating the time to market (and reducing the financial resources required) of products manufactured using recombinant DNA and living microbes, cells, transgenic plants or transgenic mammals. However, when an entirely new upstream platform or downstream unit operation is needed, there are few books that will quickly provide the depth of industry-relevant background. Downstream Industrial Biotechnology can fill this void as an advanced desk reference. This volume includes relevant biology, protein purification and engineering literature with abundant process examples provide by industry subject matter experts (SMEs) and academic scholars. This desk reference will also be useful for advanced biomanufacturing students and professionals to quickly gain in depth knowledge on how to design processes (and facilities) capable of being licensed to manufacture enzymes, biopharmaceutical intermediates, human and veterinary biopharmaceuticals or vaccines. The opportunity is yours to leverage the combined knowledge from scores of industry professionals from around the world who have contributed to Downstream Industrial Biotechnology to reduce the time and cost to deliver engineered proteins, biomolecules and cost-effective biologics to the market and especially to millions of patients worldwide.

Professor Michael C. Flickinger, Editor

Golden LEAF Biomanufacturing Training and Education Center (BTEC)

Department of Chemical and Biomolecular Engineering

North Carolina State University

Raleigh, North Carolina, 27695-7928, USA

Contributors

Muhammad Aasim Downstream Bioprocessing Laboratory, School of Engineering and Science, Jacobs University, Bremen, Germany

Oscar Aguilar Centro de Biotecnologa Tecnologico de Monterrey, Monterrey, Mexico

Mattias Ahnfelt GE Healthcare Bio-Sciences AB, Uppsala, Sweden

Hazel Aranha GAEA Resources Inc., Northport, New York, USA

Claude Artois University of Surrey, Guildford, Surrey, United Kingdom; SmithKline Beecham Biologicals, Rixensart, Belgium

Hans Axelsson Alfa Laval AB, Tumba, Sweden

Diana C.S. Azevedo Federal University of Ceara, Fortaleza-CE, Brazil

H.S.C. Barbosa Center of Chemistry, University of Minho, Campus de Gualtar, Braga, Portugal

Sonja Berensmeier Technische Universitat Munchen, Institute of Biochemical Engineering, Garching, Germany

Joseph Bertolini CSL Bioplasma, Broadmeadows, Victoria, Australia

Darcy Birse Fast Trak Biopharma Services, GE Healthcare, Piscataway, New Jersey, USA

Eggert Brekkan GE Healthcare Bio-Sciences AB, Uppsala, Sweden

Phil J. Bremer University of Otago, Dunedin, New Zealand

Kurt Brorson Office of Biotech Products, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, USA

Kurt Brorson Office of Pharmaceutical Science, Center for Drug Evaluation and Research, United States Food and Drug Administration

Thierry Burnouf Human Protein Process Sciences, Lille, France

Trent Carrier Invitrogen, part of Life Technologies, Grand Island, New York, USA

David Clark Centocor R&D, Spring House, Pennsylvania, USA

Efrem Curcio University of Calabria, Arcavacata di Rende (CS), Italy

Jean Didelez University of Surrey, Guildford, Surrey, United Kingdom; SmithKline Beecham Biologicals, Rixensart, Belgium

Gianluca Di Profio Institute on Membrane Technology (ITM-CNR), c/o University of Calabria, Arcavacata di Rende (CS), Italy; University of Calabria, Arcavacata di Rende (CS), Italy

Dennis Dobie Fluor Daniel, Marlton, New Jersey, USA

Ed Domanico Tri-Clover, Valencia, California, USA

Enrico Drioli Institute on Membrane Technology ITM-CNR, At University of Calabria, Rende, Italy

Zhiwu Fang Amgen Inc., Systems Informatics, Thousand Oaks, California, USA

Patrick Florent University of Surrey, Guildford, Surrey, United Kingdom; SmithKline Beecham Biologicals, Rixensart, Belgium

Matthias Franzreb Karlsruhe Institute of Technology, Institute for Functional Interfaces, Eggenstein-Leopoldshafen, Germany

Pete Gagnon Validated Biosystems, San Clemente, California, USA

F.A.P. Garcia University of Coimbra, Coimbra, Portugal

Tom Gervais Centocor R&D Spring House, Pennsylvania, USA

Iraj Ghazi The Ohio State University, Columbus, Ohio, USA

Siddartha Ghose Aston University, Birmingham, United Kingdom

Guy Godeau University of Surrey, Guildford, Surrey, United Kingdom; SmithKline Beecham Biologicals, Rixensart, Belgium

Susanne Graslund Structural Genomics Consortium, Karolinska Institutet, Stockholm, Sweden

Tingyue Gu Ohio University, Athens, Ohio, USA

Martin Hammarstrom Structural Genomics Consortium, Karolinska Institutet, Stockholm, Sweden

Richard Hassett Invitrogen, part of Life Technologies, Grand Island, New York, USA

Eva Heldin GE Healthcare Bio-Sciences AB, Uppsala, Sweden

Nathaniel G. Hentz, PhD North Carolina State University, Golden LEAF Biomanufacturing Training and Education Center, Raleigh, North Carolina, USA

Birgit Hickstein Clausthal University of Technology, Institute of Chemical Process Engineering, Clausthal-Zellerfeld, Germany

Timothy John Hobley Technical University of Denmark, Systems of Biology, Lyngby, Denmark

Tony Hunt Advanced Minerals Corporation, Santa Barbara, California, USA

Omkar Joshi Bayer HealthCare LLC, Berkeley, California, USA

Varsha S. Joshi Chemical Engineering Department, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, India

Adalberto Pessoa Jr. School of Pharmaceutical Sciences, University of Sao Paulo, Brazil

Amaro G. Barreto Jr. Escola de Quimica, Universidade Federal do Rio de Janeiro, Rio de Janeiro-RJ,Brazil

Ivanildo J. Silva Jr. Federal University of Ceara, Fortaleza-CE, Brazil

Beth H. Junker Bioprocess R&D Merck Research Laboratories, Rahway, New Jersey, USA

Manohar Kalyanpur Consultant, Bioseparations & Pharmaceutical Validation, Plaisir, France

Ingo Kampen Technische Universitat, Institute for Particle Technology, Braunschweig, Germany

Mansoor A. Khan Office of Pharmaceutical Science, Center for Drug Evaluation and Research, United States Food and Drug Administration

Alexandros Koulouris Intelligen Europe, Thermi, Greece

Maria-Regina Kula Heinrich Heine University Dusseldorf, Julich, Germany

Ingo Kampen Arno Kwade Technische Universitat, Institute for Particle Technology, Braunschweig, Germany

Per Kaarsnas Institute of Biology and Chemical Engineering, Malardalens hogskola, Eskilstuna, Sweden

Marcelo Fernandez Lahore Downstream Bioprocessing Laboratory, School of Engineering and Science, Jacobs University, Bremen, Germany

Philippe Lam Pharmaceutical Development Genentech, Inc., South San Francisco, California, USA

Chung Lim Law The University of Nottingham, Malaysia Campus, Selangor, Malaysia

Jinsong Liu Product Development, Abraxis BioScience, Melrose Park, Illinois, USA

Scott Lute Office of Biotech Products, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, USA

Perola O. Magalhaes University of Brasilia, Brasilia, DF, Brazil

Robert Z. Maigetter Centocor R&D, Spring House, Pennsylvania, USA

J.C. Marcos Center of Chemistry, University of Minho, Campus de Gualtar, Braga, Portugal

Joseph McGuire Oregon State University, Corvallis, Oregon, USA

George Miesegaes Office of Biotech Products, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, USA

Jamie Moore Pharmaceutical Development Genentech, Inc., South San Francisco, California, USA

Manuel Mota IBB, Centro de Eng. Biologica, University of Minho, Portugal

Arun S. Mujumdar National University of Singapore, Singapore

P.T. Noble Fluor Daniel GmbH, Wiesbaden, Germany

Jeffery N. Odum CPIP Biotech Sector Lead & Director of Operations Integrated Project Services

Jeffery Odum IPS, Morrisville (RTP), North Carolina, USA

Victor Papavasileiou Intelligen Europe, Leiden, The Netherlands

Jun T. Park Office of Pharmaceutical Science, Center for Drug Evaluation and Research, United States Food and Drug Administration

Steve Peppers Invitrogen, part of Life Technologies, Grand Island, New York, USA

Demetri Petrides Intelligen, Inc., Scotch Plains, New Jersey, USA

Urs Alexander Peuker TU Bergakademie Freiberg, Institute for Mechanical Process Engineering and Mineral Processing, Freiberg, Germany

John Pieracci Biogen Idec, San Diego, California, USA

Tom Piombino Integrated Project Services, Inc., Lafayette Hill, Pennsylvania, USA

Tina Pitarresi Fast Trak Biopharma Services, GE Healthcare, Piscataway, New Jersey, USA

Mirjana Radosevich Human Protein Process Sciences, Lille, France

Anurag S. Rathore Chemical Engineering Department, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, India

Erik K. Read Office of Pharmaceutical Science, Center for Drug Evaluation and Research, United States Food and Drug Administration

James J. Reilly Laureate Pharma, Inc., Princeton, New Jersey, USA

Robert van Reis Genentech, Inc., South San Francisco, California, USA

Craig Robinson GE Healthcare, Westborough, Massachusetts, USA

Carl A. Rockburne The Rockburne Group, Atlanta, Georgia, USA

Gustav Rodrigo GE Healthcare Bio-Sciences AB, Uppsala, Sweden

Cesar C. Santana School of Chemical Engineering, State University of Campinas, Campinas-SP, Brazil

Maria Schafer TU Bergakademie Freiberg, Institute for Mechanical Process Engineering and Mineral Processing, Freiberg, Germany

Catherine H. Schein Sealy Center for Structural Biology and Molecular Biophysics, Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas, USA

Richard Brent Seale University of Otago, Dunedin, New Zealand

Klaus Selber Heinrich Heine University Dusseldorf, Julich, Germany

Rakhi B. Shah Office of Pharmaceutical Science, Center for Drug Evaluation and Research, United States Food and Drug Administration

Bryan Shingle Centocor R&D Spring House, Pennsylvania, USA

Charles Siletti Intelligen, Inc., Mt. Laurel, New Jersey, USA

Eduardo V. Soares Bioengineering Laboratory, Superior Institute of Engineering from Porto Polytechnic Institute, Porto, Portugal; IBB-Institute for Biotechnology and Bioengineering, Centre for Biological Engineering, Universidade do Minho, Braga, Portugal

Gail Sofer GE Healthcare, Piscataway, New Jersey, USA

Bob Stover Tri-Clover, Valencia, California, USA

Jorg Thommes Biogen Idec, San Diego, California, USA

Owen Thomas University of Birmingham, Biochemical Engineering, Birmingham, United Kingdom

Claudio Thomasin Centocor R&D, Spring House, Pennsylvania, USA

Michiel E. Ultee Laureate Pharma, Inc., Princeton, New Jersey, USA

Greg Van Slyke Invitrogen, part of Life Technologies, Grand Island, New York, USA

Rami Reddy Vennapusa Downstream Bioprocessing Laboratory, School of Engineering and Science, Jacobs University, Bremen, Germany

Gary Walsh Industrial Biochemistry and Materials Surface Sciences Institute, University of Limerick, Limerick City, Ireland

Dave A. Wareheim Centocor R&D Spring House, Pennsylvania, USA

Sandy Weinberg Clayton State University, Atlanta, Georgia, USA

Christian Wood Centocor R&D Spring House, Pennsylvania, USA

David W. Wood The Ohio State University, Columbus, Ohio, USA

Alexander Yelshin Polotsk State University,~Novopolotsk, Belarus

Inna Yelshina Polotsk State University, Novopolotsk, Belarus

Jonathan Yourkin GE Instruments, Boulder, Colorado

David (Xiaojian) Zhao Invitrogen, part of Life Technologies, Grand Island, New York, USA

Andrew L. Zydney The Pennsylvania State University, University Park, Pennsylvania, USA

Part I

Introduction

Introduction

Downstream biomanufacturing processes increase product concentration and purity, while decreasing process volume. Therefore, decreasing process volume without loss of product is essential to increase product purity, while at the same time eliminating product contaminants. The biochemistry of different products (peptides, proteins, hormones, low-molecular-weight metabolic intermediates, complex antigens etc.), all of which are liable to degradation, dictates that different separation methods be used to isolate and purify these products from contaminating biomolecules produced by the upstream process. Optimal downstream product yield is the yield of recovered product in the appropriate final biologically active form and purity. Purified but inactive product is a contaminant, reduces overall process yield, and may have serious consequences on clinical safety and efficacy. That is why downstream process design has the greatest impact on the overall biomanufacturing cost.

As product purity increases, more product can be lost to inactivation, nonspecific binding to equipment surfaces, binding to membranes, and chromatography media or by precipitation, thus decreasing the recovery of product. Because of these potential losses, each additional separation step may reduce overall yield. Therefore, downstream separation scientists and engineers are continually seeking to eliminate or combine unit operations to minimize the number of process steps in order to maximize product recovery at a specified concentration and purity.

Section II of Downstream Industrial Biotechnology includes detailed methods used for the initial steps of cell separation, cell disruption (for intracellular products), filter aids and adsorbents for rapid protein capture and initial volume reduction. Each of these steps is critically affected by upstream process design (volume, product concentration, and contaminants derived from the growth media or host cells), which impacts every subsequent step of downstream product recovery and purification. In particular, cell separation and cell disruption methods can have a dramatic effect on contributing (or minimizing) contaminants such as nucleic acids, host cell proteins, cell membrane fragments or pyrogenic lipopolysaccharides that need to be removed from the final product in subsequent separation steps.

Although each upstream process decision impacts downstream product recovery and purification, not all contaminants come from upstream operations. In some cases contaminants can also be generated by downstream operations, as inactivated product (due to heating, proteolysis, photoinactivation or precipitation), bioburden or microbial contamination introduced during downstream operations (from the environment, water, operations staff etc.) or contaminants derived from materials in direct contact with the product (extractable, leachable contaminants).

The downstream steps described in Section III are optimized by absorbent surface area, selectivity, binding capacity, and degree of volume reduction to purify product in the concentration range needed for each subsequent step to meet overall criteria of scale, stability, purity, and potency. Therefore, close integration of the characteristics of the upstream biological system that produces the product with the engineering and optimal performance of the downstream product separation, concentration. and purification operations are essential. This means that separation engineers, bioseparation and bioanalytical scientists, and manufacturing operations staff with broad expertise in working with labile biological molecules all need to work and communicate effectively as a team to design a downstream process that can be scaled from the laboratory bench and transferred to the manufacturing scale. It also means that downstream process scientists must continually provide feedback information to upstream process engineers and scientists to minimize the impact of upstream changes (cell line changes, media composition changes, the addition of antifoam, degradation of product during in-process storage or holds) on downstream separation operations. Therefore, the companion volumes of Upstream Industrial Biotechnology should also be consulted when designing a downstream process.

Each downstream step requires process development and optimization (for purity, overall yield) because of the complexity of the structure of the biological molecules being purified and the complexity of contaminants. Section III also includes approaches for scale down of purification operations. Each downstream step is expensive to optimize at the pilot or manufacturing scale. This expense is not only due to the scale of the equipment and expense of the separation media, but also because of the large quantity of valuable product needed to carry out optimization studies at scale.

Downstream operations require specialized equipment designed for separation of proteins, peptides, virus, particulate antigens or low-molecular-weight biomolecules while minimizing product degradation. Sections IV and V focus on large scale equipment design and fluid transfer systems, and describe in detail many types of industrial bioseparation equipment. Of particular concern for products derived from mammalian cell lines are effective methods for virus inactivation and viral filtration that can be validated with model virus challenge. These methods are described in section V.

Not only do the upstream and downstream processes need to be designed to meet cGMPs and be capable of being licensed, but the facility used to carry out the process also must be designed so that it can be licensed. Section VI and VII of Downstream address facility design, facility validation, clean-in-place (CIP) and sterilization-in-place (SIP) methods. A major advance in facility design for downstream processes is the growing impact of single use (SU) disposable downstream materials and this is described in Section VI.

The overall goal of all downstream operations is not only to purify bulk product for formulation, but to achieve regulatory compliance and licensure so that final formulated and filled product can be released to consumers, physicians or patients. Section VII describes how Process Analytical Technology (PAT), bioburden testing and Quality by Design (QbD) impact downstream process design and contribute to regulatory compliance both for the USFDA and European regulatory agencies.

Chapter 1

Bioprocess Design, Computer-Aided

Victor Papavasileiou

Intelligen Europe, Leiden, The Netherlands

Charles Siletti

Intelligen, Inc., Mt. Laurel, New Jersey

Alexandros Koulouris

Intelligen Europe, Thermi, Greece

Demetri Petrides

Intelligen, Inc., Scotch Plains, New Jersey

1.1 Introduction

Bioprocess design is the conceptual work done prior to commercialization of a biological product. Given information on the potential market demand for a new product, bioprocess design endeavors to answer the following questions: What are the required amounts of raw materials and utilities for manufacturing a certain amount of product per year? What is the required size of process equipment and supporting utilities? Can the product be manufactured in an existing facility or is a new plant required? What is the total capital investment for a new facility? What is the manufacturing cost? How long does a single batch take? What is the minimum time between consecutive batches? During the course of a batch, what is the demand for various resources (e.g. raw materials, labor, and utilities)? Which process steps or resources are the likely production bottlenecks? What process and equipment changes can increase throughput? What is the environmental impact of the process? Which design is the “best” among several plausible alternatives?