Cells and Culture: Proceedings of the 20th ESACT Meeting, Dresden, Germany, June 17-20, 2007 (ESACT Proceedings, 4)

It has been predicted that nearly half of all human proteins are glycosylated indicating the significance of glycoproteins in human health and disease. For example, the glycans attached to proteins have emerged as important biomarkers in the diagnosis of diseases such as cancers and play a significant role in how pathogenic viruses gain entry into human cells. The study of glycoproteins has now become a truly proteomic science. In the last few years, technology developments including in silico methods, high throughput separation and detection techniques have accelerated the characterization of glycoproteins in cells and tissues. Glyco-engineering coupled to rapid recombinant protein production has facilitated the determination of glycoprotein structures key to exploring and exploiting their functional roles. Each chapter in this volume is written by experts in the field and together provide a review of the state of the art in the emerging field of glycoproteomics. Front Matter....Pages i-liv Front Matter....Pages 1-1 Targeted Gene Knockdown Effects on Recombinant Protein Secretion in CHO Cells....Pages 3-8 RNA Silencing Suppressors Boost the Production of Recombinant Proteins and Viruses....Pages 9-12 Towards the Use of CHO Produced Recombinant Extracellular Matrix Proteins as Bioactive Elements in a 3-D Scaffold for Tissue Engineering....Pages 13-18 Transient Gene Expression in Chinese Hamster Ovary Cells at Low Temperature – The Effects of Cold-Induced Proteins and an mRNA Regulatory Element....Pages 19-23 Single-Cell Approach in Influenza Vaccine Production: Apoptosis and Virus Protein Production....Pages 25-28 Chondrogenic Differentiation of Human Mesenchymal Stem Cells During Multiple Subcultivation....Pages 29-34 Cell XpressTM-Assisted Analysis of Clone Stability in Recombinant Chinese Hamster Ovary Cells....Pages 35-39 An Evaluation of the Intrinsic IgG Production Capabilities of Different Chinese Hamster Ovary Parental Cell Lines....Pages 41-44 Cell XpressTM Technology Facilitates High-Producing Chinese Hamster Ovary Cell Line Generation Using Glutamine Synthetase Gene Expression System....Pages 45-48 CHO-DG44 Cell Line Development by FLP-Targeting – High Level Glycoprotein Expression with Significantly Decreased Time Lines....Pages 49-55 Transgene Copy Number – Impact on Clone Performance....Pages 57-62 Single Use Bioreactors: Expressing Protein in Mammalian Cell Suspension....Pages 63-69 Front Matter....Pages 71-71 Gene Modified Hematopoietic Stem Cells for the Treatment of Primary Immunodeficiency Diseases....Pages 73-78 An Avian Cell Line for Production of Highly Attenuated Viral Vectors....Pages 79-88 Microelectronic Cellular Vitality Monitoring....Pages 89-93 IFN-γ Glycosylation Macroheterogeneity, Energetic Cell Status and Medium Composition During CHO Cell Cultures....Pages 95-99 Modelling of Neural Metabolism Using 13 C-NMR Spectroscopy and Metabolic Flux Analysis....Pages 101-106 Influence of Glucose and Glutamine Concentration on Metabolism of rCHO Cells....Pages 107-111 Front Matter....Pages 113-113 Delivery of Biomolecules with Non-Viral Vectors....Pages 115-119 Circumventing the “Pay Now or Pay Later” Dilemma: Strategies for Achieving Process Development with Speed and Long-Term Potential....Pages 121-128 Front Matter....Pages 113-113 Recombinant Human Antibody Therapeutics: Supply Strategies for Early and Clinical Development from CHO Cells....Pages 129-136 Automated Screening of High Producer HEK293F Clones and Analysis of Post-Translational Modifications of Secreted Proteins....Pages 137-144 Transcriptomic and Proteomic Analysis of Antibody Producing NS0 Cells Cultivated at Different Cell Densities in Perfusion Culture....Pages 145-149 New Disposable Fixed-Bed Bioreactor for Cell Culture and Virus Production Based on a Proprietary Agitation and Aeration System....Pages 151-155 Transcriptomic Analysis of Antibody Producing NS0 Cell Line Under Hypothermic and Hypoxic Conditions....Pages 157-160 Semi-Continuous Cultures as a Tool for Cell Line Characterization During Process Development....Pages 161-164 Behaviour of GS-CHO Cell Lines in a Selection Strategy....Pages 165-169 3D Cultures: Effect on the Hepatocytes Functionality....Pages 171-176 Differential Protein Expression Induced by c-Myc Over-Expression: Proteomic Analysis of a CHO Cell Line with Increased Proliferation Capacity....Pages 177-181 Development of Pilot-Scale Orbital Shake Bioreactors: Ideal for Cost-Effective and Efficient Transient Gene Expression....Pages 183-185 Helical Tracks in Shaken Cylindrical Bioreactors Improve Oxygen Transfer and Increase Maximum Cell Density Obtainable for Suspension Cultures of Mammalian Cells....Pages 187-191 Dynamic Optimisation of CHO-IFNγ Cell Culture Fed-Batch Time-Profile....Pages 193-199 Long-Term 3D-Culture of HEP G2 Cell Line on Macroporous Ceramic Carriers....Pages 201-204 An Integrated Production Process for Human Growth Hormone....Pages 205-208 Efficient Production of Human Monoclonal Antibodies by an Improved Fructose-Based Human Cell Culture....Pages 209-212 Coupling Between Cell Kinetics and CFD to Establish Physio-Hydrodynamic Correlations in Various Stirred Culture Systems....Pages 213-217 Confocal Microscopy Observation of CHO Cells Cultivated in Presence of Fluorescent – Labelled Rapeseed Peptides....Pages 219-222 Growth and Production Characteristics of Four Mammalian Cell Lines on a Cost-Effective Serum-Free Medium....Pages 223-227 A Serum-Free, Transient Transfection System for Enhancing Production of Recombinant Antibodies in Mammalian Cells....Pages 229-232 CFD Study of the Fluid and Particle Dynamics in a Spin-Filter Perfusion Bioreactor....Pages 233-238 Front Matter....Pages 113-113 High-Yielding CHO Cell Pools for Rapid Production of Recombinant Antibodies....Pages 239-244 Increasing Upstream Process Development Efficiency by Implementing Platform Glutamine Synthetase Cell Culture Processes....Pages 245-251 Implementation of High Throughput Systems for Media and Process Development....Pages 253-255 Comparison of Cell Culture Methods for Obtaining of rHU-EPO to Large Scale....Pages 257-260 Optimization and Comparison of Different DNA Methyl Transferase and Histone Deacetylase Inhibitors for Enhancing Transient Protein Expression....Pages 261-264 Proteomic Characterisation of a Glucose-Limited CHO Perfusion Process–Analysis of Metabolic Changes and Increase in Productivity....Pages 265-269 Evaluation of Alternative Signal Sequences....Pages 271-274 Process Development for the GMP Production of N-Acetylgalactosamine-6-Sulfate Sulfatase (GALNS) Expressed by CHO Cells....Pages 275-279 Improvement of a CHO Fed-Batch Process by Fortifying with Plant Peptones....Pages 281-284 O -Glycans on Recombinant MUC1 Produced in CHO K1 Cells Become Less Sialylated with Increased Protein Productivity, as Determined by LC-ESI MS....Pages 285-288 A Multiple Minibioreactor Platform for Parallel and Automated Mammalian Cell Culture....Pages 289-293 Cultivation of Adherent-Dependent Animal Cells on Microcarriers in a New Disposable Reactor....Pages 295-299 CHO Cells Cultivation and Antibodies Production in a New Disposable Bioreactor Based on Magnetic Driven Centrifugal Pump....Pages 301-305 Stability and Productivity of CHO Pools with Respect to Culture Age, Cryopreservation and 20 L Bioreactor Cultivation....Pages 307-311 In Vitro Disassembly and Reassembly of Triple-Layered Rotavirus-Like Particles....Pages 313-318 Influence of Culture Conditions on Insect Cell Growth and Protein Production – Comparison of Wave Bioreactor and Shake Flasks....Pages 319-324 Process Intensification Based on Nano-Structured Carbon Carrier Materials and Disposable Devices....Pages 325-330 Accelerating Fed-Batch Process Development Using Rationally Designed Feed Media....Pages 331-336 Development of a Robust Small-Scale Production Format that Is Predictive of Bioreactor Performance....Pages 337-342 Biomass Monitoring and CHO Cell Culture Optimization Using Capacitance Spectroscopy....Pages 343-348 Front Matter....Pages 113-113 Characterizing Physiology and Metabolism of High-Density CHO Cell Perfusion Cultures Using 2D-NMR Spectroscopy....Pages 349-357 BI HEXTM – Platform for Fast Track Generation of High Producer Cell Lines Leading to High-Titer Processes for Production of Therapeutic Proteins from Mammalian Cells....Pages 359-363 Management of Handling, Long-Term Stability and Shipping of Human T-Lymphocytes Bags for Clinical Studies....Pages 365-369 Mass Transfer in the CELL-tainer ® Disposable Bioreactor....Pages 371-373 Efficient Production of Human Monoclonal Antibodies by Combining In Vitro Immunization and Phage Display Methods....Pages 375-377 Measurement and Control of Viable Cell Density in a Mammalian Cell Bioprocessing Facility: Validation of the Software....Pages 379-384 Development, Validation, and Application of a Fully Integrated Automation System for Screening and Selection of High Yielding Production Cell Lines....Pages 385-389 Monitoring of Cell Activity: Online Oxygen Uptake Rates in Pulsed Aerated Cell Culture....Pages 391-393 Front Matter....Pages 395-395 Metabolite Analysis in Mammalian Cells: How to Generate Reliable Data Sets?....Pages 397-406 Cell-to-Cell Communication for Cell Density-Controlled Bioprocesses....Pages 407-412 Metabolome Analysis in Mammalian Cells: Development and Application of a Sampling Technique....Pages 413-420 Differential Expression Profiling of Industrially Relevant CHO Cell Phenotypes Using a Proprietary CHO-Specific Microarray and Proteomics Technology Platforms....Pages 421-423 Towards a Systems-Level Understanding of Increased Specific Productivity in Proliferation Arrested Myeloma NS0 Cells....Pages 425-428 Proteomic Analysis of Influenza a Virus Infected Mammalian Cells by 2D-DIGE....Pages 429-432 Involvement of SRC- and MAP Kinase-Signalings in the Effect of Sericin on Cellular Proliferation and Survival....Pages 433-435 The Unfolded Protein Response and Recombinant Protein Production from In Vitro Cultured Mammalian Cells....Pages 437-440 A Scalable Process for Helper-Dependent Adenoviral Vector Production Using PEI-Derived Transfection Strategy in Suspension Culture....Pages 441-445 Functional Analysis of ER Stress Pathway Genes for Apoptosis of NS/0 Cell Line Using RNAi Methods....Pages 447-451 Identification of New Protein Substrate Candidates of Transglutaminase in Rat Liver Extracts: Use of 5-(Biotinamido) Pentylamine as a Probe....Pages 453-458 Metabolic Flux Distributions of Adherently Growing MDCK Cells in Different Media....Pages 459-462 Front Matter....Pages 395-395 Expression of Dermal Extracellular Matrix Proteins in a Newly Developed Full-Thickness Skin Model....Pages 463-467 Development of Preparations with Antiviral and Immunostimutory Effects from Extracts of Green Parts of Coniferous Plants....Pages 469-473 Serum-Free Transfection of CHO Cells with Chemically Defined Transfection Systems to Generate Transient and Stable Cell Lines....Pages 475-478 “ Glycomics” : Development and Characterization of Glycan-Based Biotechnological Products....Pages 479-480 Development of a Cell-Culture-Based Platform for Viral Vaccine Production....Pages 481-484 Screening of Natural Compounds Affecting Type I Interferon Signalling....Pages 485-487 Front Matter....Pages 489-489 Therapeutic Proteins from Transgenic Cows’ Milk....Pages 491-496 Development of Edible Plant Vaccines....Pages 497-501 Human Cell Lines for Production of Biopharmaceuticals....Pages 503-511 Concentration and Purification of Densonucleosis Virus by Tangential Membrane Filtration and by Ion Exchange Membranes....Pages 513-516 Effect of ManNAc and its Derivatives on Glycosylation to Proteins Produced by Mammalian Cell Culture....Pages 517-519 Purification of a Chimeric Simian – Human Immunodeficiency Virus-Like Nanoparticle from HEK293 Cell Culture....Pages 521-527 Effects of Plant Peptones Supplemented Medium on CHO Cells....Pages 529-532 Synthetic Low Density Lipoprotein a Novel Biomimetic Lipid Supplement for Serum Free Tissue Culture....Pages 533-537 Front Matter....Pages 539-539 Scale-Down Approach for Animal-Free Polio Vaccine Production....Pages 541-550 “Novel Scaffolds for Tissue Engineering – Nano-Structured Surfaces Promote Selective Cell Attachment and Cell Differentiation”....Pages 551-557 Unconventional Experimental Concepts Enabling High Speed High Performance Media/Process Development....Pages 559-565 A Study on Bioscaffolds of Polysialic Acid and β-Glucan for Cell Culture and Tissue Engineering Applications....Pages 567-572 A Preliminary Study on Spider Silk as Biomaterial for Peripheral Nerve Regeneration....Pages 573-578 Investigation of the Effect of Mechanical Strain on the Osteogenic Differentiation of Mesenchymal Stem Cells....Pages 579-589 Front Matter....Pages 539-539 Biofunctional Polymer-Mineral Composites as Scaffolds for Bone Tissue Engineering....Pages 591-597 New Water-Soluble Polymers for Construction of Biofunctionalized Scaffolds for Bone Tissue Engineering: Synthesis and Adsorption Study....Pages 599-605 Selection of High-Producing Cells Via Cell Sorting Using an Affinity Matrix....Pages 607-611 The Effects of Medium Supplement on High-Level Production of Recombinant Human Factor IX in CHO Cell....Pages 613-618 Case Study Large Scale Cell Culture Facility....Pages 619-620 Glycosylation of Influenza A Virus Hemagglutinin....Pages 621-623 Production of Retroviral Pseudotype Vectors in Fixed Bed Reactors for Use in Gene Therapy....Pages 625-628 Swellscreen – Rapid Baculovirus Titration Method in Microplate Format....Pages 629-633 Comparing Vero and MDCK Cells for Influenza A Virus Production in Microcarrier Systems....Pages 635-638 Expansion of Human Articular Chondrocytes on Microcarriers Enhances the Production of Cartilage Specific Matrix Components....Pages 639-643 Automated Cell Culture Handling – Characteristics and Procedures....Pages 645-645 Bioreactor Satellite Culture Experiments in the Start-Up of a Cell Culture Technical Support Lab....Pages 647-655 Advantages of Hydrodynamic Cell Separation in Industrial Cell Culture Processes....Pages 657-664 Optimization and Characterization of the Process for Large Volume Cell Banking in Bags....Pages 665-668 Development of Screening Method for IgA-Promoting Factors Derived from Food Extracts Using a Human Myeloma Cell Line....Pages 669-674 Cryopreservative Solution Using Sericin....Pages 675-678 Fructan as a Novel Effective Factor for Mammalian Cell Culture....Pages 679-682 Online Determination of Oxygen Uptake and Carbon Dioxide Production Rates in Mammalian Cell Culture Using Mass Spectrometry....Pages 683-687 Optimisation of mAb Concentration in Microcarrier Based Perfusion Compared to Batch/Fed-Batch Cultivation....Pages 689-692 Characterization of the Novel Human AGE1hn Cell Line for Production of Recombinant Proteins....Pages 693-697 Front Matter....Pages 539-539 Rapid Selection of Optimal Formulations for Divergent Clones Through Screening Chinese Hamster Ovary Media Library....Pages 699-702 Development of a Vaccine Candidate Against Heartwater....Pages 703-710 Animal Component Free T-Cell Culture....Pages 711-716 Characterization of Cholesterol-Independent GS-NS/0 Recombinant Antibody Cell Lines....Pages 717-721 Development of a Fed-Batch Process for the Production of a Recombinant Protein X in CHO-GS System – Case Study from the Cell to Reactor Process Ready for Pilot Scale Cultivation....Pages 723-725 Scaffolds for Articular Joint Tissue Engineering....Pages 727-733 Effect of Tunisian Aromatic Plant Extracts on Melanogenesis....Pages 735-738 Effects of Capsaicin on Energy Metabolism by Human Intestinal Epithelial Cell Line Caco-2....Pages 739-742 Human A1AT Production – Propagating a Newly Developed Human Cell Line in a Novel Disposable Perfusion Bioreactor....Pages 743-747 Characterization of Diffusion and Flow Relations in the Novel Membrane Based Perfusion Bioreactor....Pages 749-753 Protective Effect of Di-O-Caffeoylquinic Acid on Human-Derived Neurotypic SH-SY5Y Cells Against Alzheimer’s Disease Amyloid-Beta-Induced Toxicity....Pages 755-757 Flow Through Ceramic Foams – A Future Cell Culture Challenge....Pages 759-764 Measles Virus Production in MRC-5 Cells Grown on Microcarriers in a Stirred Bioreactor....Pages 765-769 A New System for the Enrichment of Cell Subclones Secreting High Levels of IgG Using Magnetic Cell Sorting (MACS ® Technology)....Pages 771-775 Soy Hydrolysate Optimization for Cell Culture Applications....Pages 777-783 The Effect of Bioreactor pH and Temperature on Protein Glycosylation in Perfusion Cultures of Mammalian Cells....Pages 785-788 Evaluation of Cell Growth Characteristics on Chitosan-Alginate Membranes to Assess Their Potential Application on Highly Exuding Skin Lesions and In Vivo Evaluation in Wounded Cat....Pages 789-794 Vaccine Production Utilizing the Potential of Microcarriers in Disposable Bioreactor....Pages 795-801 Characterization of an Anti-Idiotypic Antibody Blocking the Capacity of the HIV-1 Specific nMAb 2F5....Pages 803-806 Cultivation of PER.C6 ® Cells in the Novel CELL-TainerTM High-Performance Disposable Bioreactor....Pages 807-808 Front Matter....Pages 539-539 Scale-Up of a PER.C6 ® Fed-Batch Process in 50 and 250 L Hyclone Single Use Bioreactors Compared to 50 and 250 L Stainless Steel Bioreactors....Pages 809-810 Capacitance Sensor as a Robust Tool for Cell Culture Monitoring in Process Development and Manufacturing....Pages 811-817 Effect of Different Cell Culture Medium Surfactants on Cell Growth and Viability....Pages 819-822 Conventional Stirred Bioreactor Control System for Monitoring and Controlling pH and DO in a Wave Bioreactor....Pages 823-828 On-Line Monitoring of Vero Cells Cultures During the Growth and Rabies Virus Process Using Biomass Spectrometer....Pages 829-832 Inducing of Human IgE Antibodies by In Vitro Immunization....Pages 833-836 Oxygen Uptake Rate (OUR) Estimation in High Density Mammalian Cell Perfusion Cultures....Pages 837-841 Use of Yeast Derived Nutrients for Cell Culture in Serum-Free Media....Pages 843-846 A Novel Approach to the Production of Plant-Derived Hydrolysates Yields Medium Supplements with Enhanced Performance in Cell Culture Systems....Pages 847-852 Monitoring the Cell Size Distribution of Mammalian Cell Cultures Using On-Line Capacitance Measurements....Pages 853-859 Biosimilarity of Recombinant Human EPO Products from CHO Cell Lines: A Carbohydrate Structural View....Pages 861-865 Quantitative N-Glycan Mapping of Glycoprotein Therapeutics by HPAEC-PAD: Glycosylation Characteristics of Different Recombinant Human EPO Products....Pages 867-871 Regeneration of tissue to replace damaged or injured tissue is the goal of t- sue engineering. Biomaterials like polyglycolic acid, collagen and small-intestinal submuscosa provide a temporary scaffold to guide new tissue growth and or- nization. Typically, they need to be biodegradable, showing good cell atta- ment and proliferation and they should possess appropriate mechanical properties (Kim et al. , 2000). Synthetic polymers ful ll most of these requirements but lack cell-adhesion peptides on their surface to enhance cell attachment. Ce- adhesion peptides are present in ECM proteins like collagen and elastin. Thus a synthetic polymer coated with ECM proteins would result in a scaffold that mimics the natural cellular environment with enhanced cell attachment and p- liferation. The new bioactive scaffold will be made by combining a synthetic polymer coated with a layer of recombinant ECM proteins produced by CHO cells. The rst step consists of identifying polymers that give best results in terms of CHO cell attachment and growth. Classical techniques to determine biomass are inappropriate to evaluate 3-D structures. Thus a screening system based on stable GFP expressing CHO cells was used to compare the different scaffolds. Simple uorescent measurement after cell lysis allows determining cell attachment and p- liferation on synthetic polymers. Finally CHO cells producing human recombinant collagen I and elastin were generated. We showed that both proteins are expressed and secreted by CHO DG44 cells. 2 Materials and Methods 2.