Note: Intro -- Preface -- Contents -- Contributors -- Chapter 1: Cell Line Development -- 1.1 Introduction -- 1.1.1 Product Quantity -- 1.1.2 Product Quality -- 1.2 Two Components of Cell Line Development: The Gene of Interest (GOI) and the Cell -- 1.3 The Classical Approach: Random Integration of GOI -- 1.4 Stable Expression by Episomal Vectors -- 1.5 Targeted Integration and Locus Specific Gene Expression -- 1.5.1 Technologies of Targeted Integration -- 1.5.2 Double Strand Break Induced Homologous Recombination -- 1.5.3 Random Integration Competes with HR -- 1.5.4 Applications of DSB Induced Modifications -- 1.5.5 Cassette Exchange Approaches Based on Site-Specific Recombinases -- 1.5.6 Tagging and Targeting -- 1.5.7 Application of Targeted Integration and RMCE -- 1.5.8 Precautions and Challenges for Targeting into Defined Chromosomal Sites -- References -- Chapter 2: Transient Recombinant Protein Expression in Mammalian Cells -- 2.1 Introduction -- 2.2 Viral Transient Transfection -- 2.3 Non-viral Transient Transfection -- 2.4 Plasmid Vector Design -- 2.4.1 Plasmid Backbone and Size -- 2.4.2 Vector Elements -- 2.4.3 Episomal Replication -- 2.5 Transfection Principles -- 2.5.1 Calcium Phosphate Coprecipitation and Calfection -- 2.5.2 DNA Lipoplex Formation and Lipid-Based Transfection -- 2.5.3 Cationic Polymers and Transient Transfection with DNA Polyplexes -- 2.5.3.1 Polyplex Uptake, Plasmid Release and Translocation into the Nucleus -- 2.5.3.2 PEI Derivatives and Other Cationic Polymers as Alternatives to PEI -- 2.5.4 Transient Transfection by Electroporation -- 2.6 Cell Lines for Non-viral Transient Transfection -- 2.7 Cell Culture Media Conditions -- 2.7.1 Use of Productivity Enhancers -- 2.8 Process Strategies for Improved Recombinant Protein Production -- 2.8.1 Mild Hypothermia -- 2.8.2 High-Density Transfection and `Direct´ Transfection. , 2.8.3 Co-transfection Strategies -- 2.8.4 Large-Scale and Automated High-Throughput Applications -- References -- Chapter 3: Production of Antibodies in Hybridoma and Non-hybridoma Cell Lines -- 3.1 Introduction -- 3.2 Antibody Structure and Function -- 3.2.1 Antibody Genes -- 3.3 Therapeutic Antibodies -- 3.3.1 Polyclonal Antibodies -- 3.3.2 Monoclonal Antibodies -- 3.4 Recombinant Antibodies -- 3.4.1 Chimeric Antibodies -- 3.4.2 Humanised Antibodies -- 3.4.3 Human Antibodies -- 3.4.4 Next Generation of Therapeutic Antibodies: Antibody-Drug Conjugates, Bi-specific Antibodies and Antibody Fragments -- 3.4.4.1 Antibody Drug Conjugates (ADCs) -- 3.4.4.2 Bispecific Antibodies -- 3.4.4.3 Antibody Fragments -- 3.5 Role of Therapeutic Antibodies in Cancer and Rheumatoid Arthritis -- 3.5.1 Therapeutic Antibodies and Cancer -- 3.5.1.1 Rituxan -- 3.5.1.2 Herceptin -- 3.5.1.3 Avastin -- 3.5.2 Therapeutic Antibodies and Rheumatoid Arthritis -- 3.5.2.1 Remicade -- 3.5.2.2 Humira -- 3.5.2.3 Enbrel -- Conclusion -- References -- Chapter 4: Bioreactors for Mammalian Cells -- 4.1 Introduction to Cultivation Systems for Mammalian Cells -- 4.1.1 Requirements and Categorization of Cultivation Systems -- 4.1.2 Immobilization of Mammalian Cells -- 4.1.3 ``Single Use´´-Bioreactors -- 4.1.4 Process Strategies -- 4.1.5 Monitoring and Control -- 4.1.6 Parameters for Characterization of Bioreactors -- 4.2 Small-Scale Culture Systems -- 4.2.1 Static Culture Systems -- 4.2.2 Dynamic Culture Systems -- 4.3 Bioreactors for Suspension Culture -- 4.3.1 Cell Damage in Stirred and Bubble Aerated Bioreactors -- 4.3.2 Design of Suspension Bioreactors -- 4.3.2.1 Stirred Tank Bioreactors -- Vessel Design -- Impeller -- Aeration Systems -- Performance Parameters -- 4.3.2.2 Bubble Columns and Air-Lift Bioreactors -- 4.3.2.3 Wave Mixed Bioreactors. , 4.3.2.4 Devices for Cell and Product Retention -- 4.3.3 Scale-Up Considerations -- 4.4 Bioreactors for Immobilized Cells -- 4.4.1 Fixed Bed and Fluidized Bed Bioreactors -- 4.4.1.1 Fixed Bed Bioreactors -- 4.4.1.2 Fluidized Bed Bioreactors -- 4.4.2 ``Hollow-Fiber´´-Bioreactors -- 4.5 Bioreactor Concepts for Tissue Engineering -- 4.6 Considerations on Bioreactor Selection and Process Design -- References -- Chapter 5: Mass Transfer and Mixing Across the Scales in Animal Cell Culture -- 5.1 Introduction -- 5.2 Oxygen Mass Transfer -- 5.2.1 Basic Oxygen Mass Transfer Concepts and Equations -- 5.2.2 The Volumetric Mass-Transfer Coefficient, kLa -- 5.2.3 The Measurement of kLa -- 5.2.3.1 The Unsteady-State (Dynamic) Method -- 5.2.3.2 The Steady State Technique -- 5.2.4 Correlations for Calculating kLa -- 5.2.4.1 Stirred Bioreactors -- 5.2.4.2 Headspace Aeration and Shaken Bioreactors -- 5.2.4.3 Bubble Columns -- 5.3 Carbon Dioxide Stripping -- 5.3.1 The `Apparent´ Mass Transfer Coefficient Issue -- 5.3.2 CO2 Evolution Rate, CER, and Control of pCO2 -- 5.3.3 pH and Osmolality -- 5.4 Heat Transfer -- 5.5 Homogeneity Issues -- 5.6 Choice of Agitation Conditions and Agitator -- 5.6.1 Mean Specific Energy Dissipation Rate, -- 5.6.2 Hydromechanical Stress Issues Due to Agitation -- 5.7 Hydromechanical Stress from Sparging -- 5.8 Agitator and Sparger Choice -- 5.9 Scale-up and Ultra Scale-Down Issues -- Conclusions -- References -- Chapter 6: Hydrodynamic Damage to Animal Cells -- 6.1 Introduction -- 6.2 Hydrodynamic Forces Acting on Cells -- 6.3 Experimental Studies Attempting to Quantify Cell Damage -- 6.4 Cell Damage from Sparging -- 6.5 Experimental Sublethal Effect of Hydrodynamic Stress -- Conclusion and Future Directions -- References -- Chapter 7: Monitoring of Cell Culture -- 7.1 Introduction -- 7.2 Monitoring Principles -- 7.3 Historical Perspective. , Scope of This Article -- 7.4 Parameters and Technologies: Viable Cell Density, Total Cell Density and Cell Viability -- 7.4.1 Dye-Based Methods for Monitoring Cell Density and Viability -- 7.4.1.1 Trypan Blue Dye Exclusion -- Hemoyctometer -- Automated Trypan Blue Based Cell Counting -- Additional, general aspects of trypan blue based cell counting methods -- 7.4.1.2 Fluorescence Based Cell Density and Viability Determination via Flow Cytometry -- 7.4.2 Non-dye-based Methods for Monitoring Cell Density and Viability -- 7.4.2.1 Impedance (Electrical Resistance) Measurement -- 7.4.2.2 Capacitance Measurement -- 7.4.2.3 Electromagnetic Spectroscopic Measurements (NIR, MIR, Raman) -- Near Infrared Spectroscopy (NIRS) -- Mid Infrared Spectroscopy (MIRS) -- Raman Spectroscopy -- General advantages/disadvantages of electromagnetic spectroscopy methods for cell density and viability determination -- 7.4.2.4 In Situ Microscopy -- 7.5 Parameters and Technologies: Metabolic Parameters and Recombinant Products -- 7.5.1 Automated Analyzers (Substrate, Metabolite and Product Monitoring) -- 7.5.2 Spectroscopic Methods (Substrate, Metabolite and Product Monitoring) -- 7.5.2.1 Near Infrared Spectroscopy (NIRS) -- 7.5.2.2 Mid Infrared Spectroscopy (MIRS) -- 7.5.2.3 Raman Spectroscopy -- 7.5.3 Automated Systems for Product-Quantification -- 7.5.3.1 Surface Plasmon Resonance (Biacore Systems) -- 7.5.3.2 Bio-Layer Interferometry (Octet Systems) -- 7.5.3.3 Microfluidic Gel Electrophoresis (Caliper LapChip Systems) -- 7.6 Parameters and Technologies: Monitoring Cell Stress and Apoptosis -- 7.6.1 Flow Cytometry (FC) -- 7.6.2 Microplate/Multiwell Plate-Reader -- 7.6.3 Mass Spectrometry -- 7.6.4 Dielectrophoresis -- References -- Chapter 8: Serum and Protein Free Media -- 8.1 Introduction -- 8.2 The Advantages and Disadvantages of Serum -- 8.3 Serum-Free Media. , 8.4 Basal Media -- 8.5 Approaches for the Development of Serum-Free Media -- 8.5.1 Top-Down Approach for Serum Replacement -- 8.5.2 Bottom-Up Approach for Serum Replacement -- 8.6 A Statistical Approach to Serum-Free Media Development -- 8.7 Mitogenic Components Needed to Replace Serum -- 8.7.1 Peptide Hydrolysates -- 8.7.2 Insulin and Insulin-Like Growth Factor -- 8.7.3 Epithelial Growth Factor (EGF) -- 8.8 Transferrin: A Carrier Protein -- 8.9 Attachment Factors -- Conclusion -- References -- Chapter 9: Glycosylation in Cell Culture -- 9.1 Introduction -- 9.2 Glycosylation Structures -- 9.3 Cell Type Specific Glycosylation -- 9.4 Culture Parameters Affecting Glycosylation -- 9.5 Glycosylation Engineering and Modification of Glycan Structure -- Conclusion -- References -- Chapter 10: Modelling of Mammalian Cell Cultures -- 10.1 Scope of Bio-pharmaceutical Industry and Challenges -- 10.2 Critical Review of Mathematical Models of Biological System -- 10.3 Classifications -- 10.3.1 Models Based on Classification of Bioprocesses -- 10.3.2 Classification of the Different Forms of Mathematical (Nonlinear) Models -- 10.3.3 Black-Box, Grey-Box and White-Box Models -- 10.4 Cells, Cell Characteristics and Cell Lines -- 10.5 FDA PAT Initiative -- 10.6 Tools of Modelling -- 10.6.1 Screening of State Variables for Explanatory Correlation with Growth and Productivity -- 10.6.2 Raman Spectrophotometry: Determination of State Variables -- 10.6.3 Flow Cytometry for Determination of Cell Cycle Phases and Organelles -- 10.6.4 Systems Biology in Computational Modelling -- 10.6.5 Regression Analysis and Estimation of Model Constants (Parameters) -- 10.6.6 Data Processing of State Variables -- 10.6.7 Design of Experiments -- 10.7 Modelling of Bioprocesses -- 10.7.1 Growth and Productivity Models -- 10.7.2 Principles Behind Model Formulation. , 10.7.3 Literature Review of Growth Models.

Note: Cover -- Title Page -- Copyright Page -- Preface -- Contents -- Contributors -- 1 Monitoring the Proliferative Capacity of Cultured Animal Cells by Cell Cycle Analysis -- 2 Use of Flow Cytometry for Monitoring Antibody Productivity and Isolating High-Secreting Hybridoma Cells -- 3 Antibody Secretion Assays Using Gel Microdrops and Flow Cytometry -- 4 High-Resolution Cell Cycle Analysis of Cell Cycle-Regulated Gene Expression -- 5 Stability of Monoclonal Antibody Production in Hybridoma Cell Culture -- 6 Flow Cytometric Studies of Osmotically Stressed and Sodium Butyrate-Treated Hybridoma Cells -- 7 Flow Cytometric Analysis of Cells Obtained from Human Bone Marrow Cultures -- 8 Measurement of Intracellular pH During the Cultivation of Hybridoma Cells in Batch and Continuous Cultures -- 9 The Relationship Between Intracellular pH and Cell Cycle in Cultured Animal Cells Using SNARF-1 Indicator -- 10 Assessment of Cell Viability in Mammalian Cell Lines -- 11 Analysis of Apoptosis by Flow Cytometry -- 12 Monitoring Βaculovirus Infection and Protein Expression in Insect Cells -- 13 Use of Protein Distribution to Analyze Budding Yeast Population Structure and Cell Cycle Progression -- 14 Cell Cycle Analysis of Microorganisms -- 15 Bacterial Characterization by Flow Cytometry -- 16 Assessment of Viability of Bacteria by Flow Cytometry -- 17 Advances in the Flow Cytometric Characterization of Plant Cells and Tissues -- Index.

Note: Cell Engineering -- Editor's page -- Copyright -- Transient Expression Cell Engineering Volume 2 -- Contents -- APPLICATION OF RECOMBINANT BACULOVIRUSES IN BIOPHARMACEUTICAL RESEARCH -- ENGINEERING POST-TRANSLATIONAL PROCESSING OF RECOMBINANT PROTEINS PRODUCED IN INSECT CELL CULTURE -- A GUIDE TO SUCCESSFUL SCALE-UP OF THE BACULOVIRUS EXPRESSION SYSTEM -- TRANSIENT GENE EXPRESSION IN MAMMALIAN CELLS BASED ON THE CALCIUM PHOSPHATE TRANSFECTION METHOD -- ADENOVIRUS VECTORS IN FUNCTIONAL GENOMICS -- ADENO-ASSOCIATED VIRUS: A PROMISING TOOL FOR GENE DELIVERY -- ALPHA VIRUS VECTORS: HIGHLY EFFICIENT SYSTEMS FOR TRANSIENT GENE EXPRESSION -- TRANSIENT EXPRESSION IN MAMMALIAN CELLS: APPLICATIONS AND PERSPECTIVES -- INDEX.