Industrial Scale Suspension Culture of Living Cells

Comprehensive and comparative coverage of all suspension cell-based production methods, containing outstanding contributions from a balanced mix of industry and academia, and edited by leading figures at one of the largest companies using industrial-scale biotechnology processing. All chapters adhere to a standardized structure and the editors have carefully avoided any overlaps in content. The following are included for each host or expression system presented: (a) organisms, cells, expression systems, products, (b) basic process design aspects, (c) basic bioreactor design aspects, (d) key factors affecting process economics, (e) regulatory aspects, (f) summary and outlook.

Autorentext
The holder of a PhD in microbiology from the University of Fribourg, Switzerland, Hans-Peter Meyer served as VP Strategic Projects Biotechnology at Lonza until his retirement in early 2014. Following three years of postdoctoral studies in Stockholm, at the University of Pennsylvania, Philadelphia, and Lehigh University, Bethlehem, USA, in 1982 he joined Prof. Armin Fiechter's team as group leader at the ETH in Zurich before starting at Lonza in Visp, Switzerland in 1986, where he held a number of positions in R&D, manufacturing, and sales & marketing. He recently joined the faculty of the University of Applied Sciences and Arts of Western Switzerland, and also remains an expert at the Commission for Technology & Innovation (CTI) of the Swiss Federal Confederation.

Diego R. Schmidhalter is head of R&T within the pharma and biotechnology custom manufacturing division at Lonza Switzerland. He holds a PhD in microbiology from the University of Fribourg, Switzerland, and carried out two years of postdoctoral studies at Genencor International, California, USA. He has held various management positions at Lonza, including head of microbial anufacturing, head of the Biopharma R&D Services business, and as head of Microbial Manufacturing Science and Technology, as well as being a member of the Biopharmaceuticals business team. Dr. Schmidhalter has over 20 years of experience in the biotechnology industry in biopharmaceuticals and biochemicals process development and manufacturing, technology transfer, scaling-up fermentations right up to the 50,000-liter scale, and within the biopharmaceuticals related regulatory environment.

Zusammenfassung

The submersed cultivation of organisms in sterile containments or fermenters has become the standard manufacturing procedure, and will remain the gold standard for some time to come. This book thus addresses submersed cell culture and fermentation and its importance for the manufacturing industry. It goes beyond expression systems and integrally investigates all those factors relevant for manufacturing using suspension cultures. In so doing, the contributions cover all industrial cultivation methods in a comprehensive and comparative manner, with most of the authors coming from the industry itself. Depending on the maturity of the technology, the chapters address in turn the expression system, basic process design, key factors affecting process economics, plant and bioreactor design, and regulatory aspects.

Inhalt

Foreword v

Preface xxi

List of Contributors xxiii

The History and Economic Relevance of Industrial Scale Suspension Culture of Living Cells 1
Hans-Peter Meyer and Diego R. Schmidhalter

1 Introduction 2

2 A Short History of Suspension Culture (Fermentation) 2

2.1 Ethanol, Organic Acids, and Solvents, the Beginning 2

2.2 Vitamins Fermentation Takes a Long Time to Develop 4

2.3 Steroids, the First Large-Scale Biocatalysis Processes 5

2.4 Antibiotics, a US-Lead Turning Point in Fermentation Technology 5

2.5 Amino Acids, a Japanese Fermentation Success Story 8

2.6 Enzymes, a European Fermentation Success Story 9

2.7 Single Cell Proteins, an Economic Flop 9

2.8 Biofuels are Controversial Story 10

2.9 Recombinant DNA Technology Based Products (Monoclonal Antibodies and Other Recombinant Proteins), Setting off an Avalanche of New Products 11

3 The Contemporary Situation 11

3.1 How Long Can the USA Keep its Leading Role? 11

3.2 China and India Become Global Forces in Fermentation 12

4 The Future of Suspension Culture 13

4.1 New Frontiers 14

4.2 Yet Uncultured Cells and Organisms? 15

5 Economic and Market Considerations 16

5.1 The Pharmaceutical Market 19

5.2 Personal Care Products 25

5.3 Chemicals, Industrial and Technical Enzymes 27

5.4 Food, Dietary Supplements (Functional Food, Nutraceuticals), and Feed Products 27

6 Conclusions 32

References 34

Part I Suspension Culture of Bacteria, Yeasts, and Filamentous Fungi 39

**1 Bacterial Suspension Cultures 41
**Patrick Sagmeister, Mohammadhadi Jazini, Joachim Klein, and Christoph Herwig

1.1 Introduction 41

1.2 Organisms, Cells, and their Products 42

1.2.1 Bacteria as Production Platform for Various Products 42

1.2.2 Historical Outline for Escherichia coli 44

1.2.3 Industrial Aspects of Bacterial Expression Systems 45

1.3 Bioprocess Design Aspects for Recombinant Products 51

1.3.1 Bacterial Cultivation Processes 51

1.3.2 Gram Negative Cell Factory: Cellular Compartments and Transport across Membranes 52

1.3.3 Industrial Strategies: Quality, Folding State, and Location of Recombinant Protein Products 53

1.3.4 Approaches towards Bioprocess Design, Optimization, and Manufacturing 55

1.3.5 Bacterial Bioprocess Design 56

1.3.5.1 Technical and Physiological Constraints for Bacterial Bioprocess Design 56

1.3.5.2 Media Design 57

1.3.5.3 Product Titer is Determined by the Biomass Concentration and the Specific Productivity qp 58

1.3.6 Industrial Production Strategy by Two-Step Cultivation 59

1.3.6.1 Batch Phase for the Accumulation of Biomass 60

1.3.6.2 Structured Approach Towards Batch Design 60

1.3.6.3 Fed-Batch Phase Process Design from Scratch 61

1.3.6.4 Induction Phase: Product Formation Characteristics 63

1.3.6.5 Process Parameters Impacting Recombinant Product Formation 64

1.3.6.6 Concept of TimeSpace Yield 65

1.4 Basic Bioreactor Design Aspects 66

1.4.1 Introduction 66

1.4.2 Vessel Design and Construction 67

1.4.3 Dimensioning 67

1.4.3.1 Materials of Construction 67

1.4.3.2 Surface Quality and Welding 69

1.4.3.3 Nozzles and Ports 70

1.4.4 Mass Transfer 70

1.4.5 Cleaning in Place 72

1.4.6 Steaming in Place 73

1.4.7 Monitoring and Control of Bioprocesses 73

1.4.7.1 Standard Instrumentation Measuring and Control of Process Parameters 73

1.4.7.2 Challenges with Bioreactor Standard Sensors 74

1.4.7.3 Advanced Bioprocess Analytics: Real-Time Monitoring of Process Variables 74

1.5 Single Use Bioreactors for Microbial Cultivation 76

1.5.1 Multi-use or Single Use? 76 1.5.2 Challenges for t...