Author: K. John Morrow, Jr. Ph.D.
CHAPTER 1
INTRODUCTION
1.1. Bioprocessing in the 21st Century
A Word of Caution
State of the Art
What Is Bioprocessing?
An Industry Committed to Mammalian Cell Expression Systems (Golden 2009, See Appendix)
A Changing Landscape
The Pipeline
Antibodies Rule the Roost
Corporate Strategies
Pressures and Opportunities
1.2. Bioprocessing Advances Are Incremental in Scope
1.3. Range of the Industry Challenge
CHAPTER 2
UPSTREAM PROCESSING
2.1. Mammalian Cell Culture Expression Systems
Historical Background
Media Development and the Evolution of Therapeutic Antibodies
Cell Line Development: Engineering for Better Performance
Developing Overproducing Cell Lines
Medarex’ Selection Procedure to Avoid Royalties
Conventional Cell Lines: CHO, NSO, PER.C6, Other Types
Chinese Hamster Ovary (CHO) Cells
NSO Cells (Nonsecreting Mouse Myeloma)
Per.C6 Cells (Golden 2009, See Appendix) (Primary Human Retinal Cells)
Human Cell Lines
Selection for Overproducers
2.2. Mammalian Cell Culture Media
Introduction to Mammalian Cell Culture Media Strategies
Elimination of Serum From Mammalian Cell Culture Media
Protein Hydrolysates
Online Sources of Information
Many Options for Cell Culture Media Optimization
2.3. Post-Translational Modifications
Glycosylation
S-Glutathionylation
Other Modifications
2.4. Other Production Alternatives
Bacteria
Yeast
Green Leafy Plants
Chlamydomonas reinhardtii
Microalgae
Insect Cells
Chesapeake PERL, Inc.
Alternatives to Baculovirus for Insect Cell Protein Synthesis
Protozoan Expression Hosts
2.5. Bioreactors: Design and Function
Computer Modeling of Bioreactor Performance
Disposable Bioreactor Components
Disposables and Rapid Response
The Rise and Fall of Large Antibody Production Facilities
CHAPTER 3
DOWNSTREAM PROCESSING
3.1. Disposables, Reusables
Disposable Technology Comes of Age
Disposable Storage Options
The Disposable Conundrum
The Future of Disposables
3.2. Purification Strategies
Fine-Tuning Contaminant Removal Through Filtration Technology
Current Polishing Strategies: The Toolbox
Single-Pass Tangential Flow Filtration (TFF)
Recombinant Polyclonal Purification Challenges
Filter Reconfiguration for Better Antibody Purification
Affinity Tag Purification (Culp 2009 and Tomaselli 2009, See Appendix)
Self-Cleaving Affinity Tag Purification
Protein A
Other Protein A Alternatives
Trends in Downstream Bioprocessing from the European Union
3.3. Monitoring Tools: HPLC-MS
CHAPTER 4
OPTIMIZING BIOPROCESS DEVELOPMENT
4.1. Time Is Money
Avoiding Revenue Loss
The Next Challenge
The Example of Insulin
Develop Processes That Will Survive Upscaling
Economic Savings From Disposables
Flexibility Generates Cost Savings
Lean, Not Necessarily Mean
CHAPTER 5
CURRENT REGULATORY STATUS OF BIOLOGICS
5.1. The Biologics Problem
European Guidelines on Biologics
Canadian Regulatory Policy for Biologics
Status of US Biosimilar Regulation
CHAPTER 6
CONCLUSIONS DRAWN WHILE ON THE ROAD AHEAD; THOUGHTS ON THE FUTURE OF BIOPROCESSING
6.1. Predicting the Future
Small, Incremental Steps Will Guide Progress in Bioprocessing
6.2. The Rise of Foreign Competition
China’s Advance
Drawbacks to China’s Advance
Chinese Regulatory Policy
United States and China: Price Competition
Will the United States Throw Away Its Advantages in Bioprocessing?
India
The European Threat
CMOs: Upsides and Downsides
6.3. Marketing Issues; Costs
Are Revolutionary Developments Possible in Bioprocessing?
Disposables in the Ascendant
Cell-Free Protein Production
6.4. Options and Caveats
Good News and Challenges Ahead
Short-Term Trends in Bioprocessing
R&D Is a Major Cost Driver
REFERENCES
APPENDIX
COMPANY INDEX WITH WEB ADDRESSES