Animal Cell Biotechnology: Methods and Protocols (Methods in Molecular Biology)

Animal Cell Biotechnology: Methods and Protocols, Third Edition constitutes a comprehensive manual of state-of-the-art and new techniques for setting up mammalian cell lines for production of biopharmaceuticals, and for optimizing critical parameters for cell culture from lab to final production. The volume is divided into five parts that reflect the processes required for different stages of production. In Part I, basic techniques for establishment of production cell lines are addressed, especially high-throughput synchronization, insect cell lines, transient gene and protein expression, DNA Profiling and Characterisation. Part II addresses tools for process and medium optimization as well as microcarrier technology while Part III covers monitoring of cell growth, viability and apoptosis, metabolic flux estimation, quenching methods as well as NMR-based techniques. Part IV details cultivation techniques, and Part V describes special applications, including vaccine production, baculovirus protein expression, chromatographic techniques for downstream as well as membrane techniques for virus separation. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Animal Cell Biotechnology: Methods and Protocols, Third Edition provides a compendium of techniques for scientists in industrial and research laboratories that use mammalian cells for biotechnology purposes. Part I. Cell Line Development and Selection   1. High-Throughput Synchronization of Mammalian Cell Cultures by Spiral Microfluidic Sorter             Wong Cheng Lee, Ali Asgar. S. Bhagat, and Chwee Teck Lim   2. Insect Cell Line Development Using Flp-Mediated Cassette Exchange Technology Joo Vidigal, Fabiana Fernandes, Ana S Coroadinha, Ana P Teixeira, and Paula M. Alves                                                 3. Scalable Transient Gene Expression in Adherent Mammalian Cells Using Polyethylenimine Lukas Fliedl and Christian Kaisermayer                                                 4. Scalable Transient Protein Expression Henry C. Chiou, Sanjay Vasu, Chao Yan Liu, Isabel Cisneros, Meredith B. Jones, and Jon Zmuda   5. DNA Profiling and Characterisation of Animal Cell Lines Glyn N. Stacey, Ed Byrne, and J. Ross Hawkins     Part II. Techniques for Process Development   6. Tools for High-Throughput Process and Medium Optimization Martin Jordan and Matthieu Stettler                                     7. Designing Media for Animal Cell Culture - CHO Cells, The Industrial Standard  Karlheinz Landauer   8. Feed Optimization in Fed-Batch Culture Ana Rita Costa, Maria Elisa Rodrigues, Mariana Henriques, Rosrio Oliveira, and Joana Azeredo                                                 9. Screening and Optimization of Chemically-Defined Media and Feeds with Integrated and Statistical Approaches Zhihua Xiao, Michelle Sabourin, Graziella Piras, and Stephen Gorfien   10. Evaluation of Solid and Porous Microcarriers for Cell Growth and Production of Recombinant Proteins         Maria Elisa Rodrigues, Ana Rita Costa, Mariana Henriques, Joana Azeredo, and Rosrio Oliveira   11. Microbioreactors and Scale-Down Models: Growth of CHO Cells Using the Pall Micro24 MicroReactor System Steve R.C. Warr     Part III. Cell Characterisation and Analysis             12. Monitoring Cell Growth, Viability and Apoptosis Michael Butler, Maureen Spearman, and Katrin Braasch   13. Metabolic Flux Estimation in Mammalian Cell Cultures Chetan T. Goudar, Richard K. Biener, James M. Piret, and Konstantin B. Konstantinov   14. Quenching Methods for the Analysis of Intracellular Metabolites Judith Wahrheit and Elmar Heinzle   15. NMR Methods for Metabolomics of Mammalian Cell Culture Bioreactors Nelly Aranibar and Michael D. Reily   16. 1H-NMR Protocol for Exometabolome Analysis of Cultured Mammalian Cells Tiago M. Duarte, Nuno Carinhas, Ana Carina Silva, Paula M. Alves, and Ana P. Teixeira     Part IV. Cultivation Techniques             17. Microfluidic Perfusion Culture Koji Hattori, Shinji Sugiura, and Toshiyuki Kanamori   18. Disposable Bioreactors for Inoculum Production and Protein Expression          Regine Eibl, Christian Lffelholz, and Dieter Eibl                                                 19. Configuration of Bioreactors Dirk E. Martens, E. J. van den End, and Mathieu Streefland                                     20. Process Analytical Technology (PAT) in Insect and Mammalian Cell Culture Processes - Dielectric Spectroscopy and Focused Beam Reflectance Measurement (FBRM) Damir Druzinec, Katja Weiss, Christiane Elseberg, Denise Salzig, Matthias Kraume, Ralf Prtner, and Peter Czermak   21. Cell Assessment by Atline Microscopy Alexander Babitzky, Patrick Lindner, and Thomas Scheper   22. Seed Train Optimization for Cell Culture Bjrn Frahm     Part V. Special Applications   23. Vaccine Production - Upstream Processing with Adherent or Suspension Cell Lines    Yvonne Genzel, Jana Rdig, Erdmann Rapp, and Udo Reichl             24. Efficient Processes for Protein Expression Using Recombinant Baculovirus Particles Heike Cremer, Ingrid Bechtold, Marion Mahnke, and Ren Assenberg   25. Chromatographic Techniques in the Downstream Processing of Proteins in Biotechnology             Ruth Freitag   26. Virus Separation Using Membranes         Tanja A. Grein, Ronald Michalsky, and Peter Czermak