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Full Description
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.
Contents
High-Throughput Synchronization of Mammalian Cell Cultures by Spiral Microfluidic Sorter.- Insect Cell Line Development Using Flp-Mediated Cassette Exchange Technology.- Scalable Transient Gene Expression in Adherent Mammalian Cells Using Polyethylenimine.- Scalable Transient Protein Expression.- DNA Profiling and Characterisation of Animal Cell Lines.- Tools for High-Throughput Process and Medium Optimization.- Designing Media for Animal Cell Culture - CHO Cells, The Industrial Standard.- Feed Optimization in Fed-Batch Culture.- Screening and Optimization of Chemically-Defined Media and Feeds with Integrated and Statistical Approaches.- Evaluation of Solid and Porous Microcarriers for Cell Growth and Production of Recombinant Proteins.- Microbioreactors and Scale-Down Models: Growth of CHO Cells Using the Pall Micro24 MicroReactor System.- Monitoring Cell Growth, Viability and Apoptosis.- Metabolic Flux Estimation in Mammalian Cell Cultures.- Quenching Methods for the Analysis of Intracellular Metabolites.- NMR Methods for Metabolomics of Mammalian Cell Culture Bioreactors.- 1H-NMR Protocol for Exometabolome Analysis of Cultured Mammalian Cells.-Microfluidic Perfusion Culture.- Disposable Bioreactors for Inoculum Production and Protein Expression.- Configuration of Bioreactors.- Process Analytical Technology (PAT) in Insect and Mammalian Cell Culture Processes - Dielectric Spectroscopy and Focused Beam Reflectance Measurement (FBRM).- Cell Assessment by Atline Microscopy.- Seed Train Optimization for Cell Culture.- Vaccine Production - Upstream Processing with Adherent or Suspension Cell Lines.- Efficient Processes for Protein Expression Using Recombinant Baculovirus Particles.- Chromatographic Techniques in the Downstream Processing of Proteins in Biotechnology.- Virus Separation Using Membranes.
