Downstream Industrial Biotechnology
Recovery and Purification

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.

Preface ix

Contributors xi

PART I INTRODUCTION 1

1 Bioprocess Design, Computer-Aided 5
Victor Papavasileiou, Charles Siletti, Alexandros Koulouris, and Demetri Petrides

PART II DOWNSTREAM RECOVERY OF CELLS AND PROTEIN CAPTURE 25

2 Cell Separation, Centrifugation 27
Hans Axelsson

3 Cell Disruption, Micromechanical Properties 49
Ingo Kampen and Arno Kwade

4 Cell Separation, Yeast Flocculation 65
Eduardo V. Soares

5 Cell Wall Disruption and Lysis 81
F. A. P. Garcia

6 Expanded Bed Chromatography, Surface Energetics of Biomass Deposition 95
Marcelo Fernandez Lahore, Oscar Aguilar, Rami Reddy Vennapusa and Muhammad Aasin

7 Filter Aids 107
Tony Hunt

8 Protein Adsorption, Expanded Bed 115
Siddartha Ghose

PART III PROCESS DEVELOPMENT IN DOWNSTREAM PURIFICATION 127

9 Scaledown of Biopharmaceutical Purification Operations 129
Anurag S. Rathore and Varsha S. Joshi

10 Adsorption in Simulated Moving Beds (SMB) 147
Cesar C. Santana, Ivanildo J. Silva Jr., Diana C. S. Azevedo, and Amaro G. Barreto Jr.

11 Adsorption of Proteins with Synthetic Materials 179
Joseph McGuire and Omkar Joshi

12 Affinity Fusions for Protein Purification 191
Susanne Gräslund and Martin Hammarström

13 Bioseparation, Magnetic Particle Adsorbents 201
Urs Alexander Peuker, Owen Thomas, Timothy John Hobley, Mathias Franzreb, Sonja Berensmeier, Maria Shäfer, and Birgit Hickstein

14 High Throughput Technologies in Bioprocess Development 221
Trent Carrier, Eva Heldin, Mattias Ahnfelt, Eggert Brekkan, Richard Hassett, Steve Peppers, Gustav Rodrigo, Greg Van Slyke, and David (Xiqaojian) Zhao

15 Large-Scale Protein Purification, Self-Cleaving Aggregation Tags 257
Iraj Ghazi and David W. Wood

16 Lipopolysaccharide, LPS removal, Depyrogenation 269
Pérola O. Magalhães and Adalberto Pessoa Jr.

17 Porous Media in Biotechnology 277
Manuel Mota, Alexander Yelshin, and Inna Yelshina

18 Protein Aggregation and Precipitation, Measurement and Control 293
Catherine H. Schein

PART IV EQUIPMENT DESIGN FOR DOWNSTREAM RECOVERY AND PROTEIN PURIFICATION 325

19 Cleaning and Sanitation in Downstream Processes 327
Gail Sofer, Craig Robinson, Joanthan Yourkin, Tina Pitarresi, and Darcy Birse

20 Clean-in-place 343
Phil J. Bremer and Richard Brent Seale

21 Large Scale Chromatography Columns, Modeling Flow Distribution 353
Zhiwu Fang

22 Pumps, Industrial 373
Bob Stover and Ed Domanico

PART V DOWNSTREAM cGMP OPERATIONS 389

23 Affinity Chromatography of Plasma Proteins 391
Mirjana Radosevich and Thierry Burnouf

24 Antibody Purification, Monoclonal and Polyclonal 405
James J. Reilly and Michiel E. Ultee

25 Chromatographic Purification of Virus Particles 415
Pete Gagnon

26 Chromatography, Hydrophobic Interactions 437
Per Karsnäs

27 Chromatography, Radar Flow 449
Tingyue Gu

28 Drying, Biological Materials 465
Chung Lim Law and Arun S. Mujumdar

29 Freeze-Drying, Pharmaceuticals 485
Jinsong Liu

30 Freezing, Biopharmaceutical 505
Philippe Lam and Jamie Moore

31 Membrane Chromatography 521
John Pieracci and Jörg Thömmes

32 Membrane Separations 545
Manohar Kalyanpur

33 Plasmid Purification 557
H .S. C. Barbosa and J. C. Marcos

34 Protein Chromatography, Manufacturing Scale 571
Joseph Bertolini

35 Protein Crystallization, Kinetics 579
Gianluca Di Profio, Efrem Curcio, and Enrico Drioli

36 Protein Purification, Aqueous Liquid Extraction 603
Maria-Regina Kula and Klaus Selber

37 Protein Ultrafiltration 617
Robert van Reis and Andrew L. Zydney

38 Virus Retentive Filters 641
George Miesegaes, Scott Lute, Hazel Aranha, and Kurt Brorson

PART VI BIOPHARMACEUTICAL FACILITY VALIDATION 655

39 Biopharmaceutical Facility Design and Validation 657
Jeffrey N. Odum

40 Closed Systems in Bioprocessing 677
Jeffrey Odum

41 Facility Design for Single Use (SU) Downstream Materials 685
Robert Z. Maigetter, Tom Piombino, Christian Wood, Tom Gervais, Claudio Thomasin, Bryan Shingle, Dave A. Wareheim, and David Clark

42 eGMPs for Production Rooms 715
Claude Arlois, Jean Didelez, Patrick Florent, and Guy Godeau

43 Heating, Ventilation, and Air Conditioning 731
Dennis Dobie

44 Sterilization-in-Place (SIP) 747
P. T. Noble

PART VII FDA cGMP REGULATORY COMPLIANCE 757

45 Pharmaceutical Bioburden Testing 759
Nathaniel G. Hentz, PhD

46 Chromatography, Industrial Scale Validation 775
Sandy Weinberg and Carl A. Rockburne

47 GMPs and GLSPs 795
Beth H. Junker

48 Quality by Design (QBD) 815
Rakhi B. Shah, Jun T. Park, Erik K. Read, Mansoor A. Khan, and Kurt Brorson

49 Regulatory Requirements, European Community 829
Gary Walsh

Index 843

DR. MICHAEL C. FLICKINGER is Associate Director for Academic Programs of the Golden LEAF Biomanufacturing Training and Education Center (BTEC) and a Professor of Chemical and Biomolecular Engineering at North Carolina State University, Raleigh.