Table of contents for Cell culture and upstream processing / edited by Michael Butler.

Bibliographic record and links to related information available from the Library of Congress catalog.

Note: Contents data are machine generated based on pre-publication provided by the publisher. Contents may have variations from the printed book or be incomplete or contain other coding.

1 Cell line development and culture strategies: future prospects to improve yields
Michael Butler
1.1 Introduction
1.2 Cell line transfection and selection
1.3 Increase in efficiency in selecting a producer cell line
1.4 Stability of gene expression
1.5 Optimization of the fermentation process
1.6 Apoptosis
1.7 Bioreactors
1.8 The capacity crunch
2 Use of DNA insulator elements and scaffold/matrix-attached regions for enhanced recombinant protein expression 
Helen Kim
2.1 Introduction 
2.2 The position effect 
2.3 Use of insulators and S/MARs can reduce the effects of heterochromatin on transgene expression 
2.4 DNA insulator elements 
2.5 The scaffold/matrix-attachment regions 
2.6 Binding proteins for DNA insulators and S/MARs 
2.7 DNA insulators or S/MARs can be incorporated into expression vectors 
2.8 DNA insulators and S/MARs act in a context-dependent manner 
2.9 Conclusion 
3 Targeted gene insertion to enhance protein production from cell lines 
Trevor N. Collingwood and Fyodor D. Urnov
3.1 Introduction 000
3.2 Identification of genomic 'hot spot' loci 
3.3 Recombinase-mediated site-specific gene insertion 
3.3.1 Cre, Flp, and_C31 recombinase systems 
3.3.2 Recombinase-mediated cassette exchange 
3.3.3 Gene insertion at native 'pseudo' recombinase sites 
3.3.4 Modification of recombinases and their target sites 
3.4 Emerging technologies for targeted gene insertion 
3.4.1 Homing endonucleases in HDR-mediated targeted gene insertion 
3.4.2 Targeted gene insertion into native loci by zinc finger, nucleasemediated, high-frequency, homologous recombination 
3.5 Perspective 
4 Recombinant IgG production from myeloma and Chinese hamster ovary cells 
Ray Field
4.1 Introduction 
4.2 The need for recombinant human antibodies 
4.3 Recombinant antibodies 
4.4 Decoupling antibody isolation and production 
4.5 Choice of host cells 
4.5.1 Chinese hamster ovary cells 
4.5.2 Rodent myeloma cells 
4.6 The glutamine synthetase system 
4.7 Cell line stability 
4.8 Bioreactor process strategies 
4.9 IgG supply during antibody development 
4.10 Strategies for cell line engineering during clinical development 
4.11 Cost of goods and intellectual property 
4.12 Recombinant human IgG production from myeloma and CHO cells 
4.12.1 Creation of CHO and NS0 cell lines expressing IgG 
4.12.2 Cell expansion, subculture and production reactor experiments 
4.12.3 Northern and western blotting 
4.12.5 Dilution cloning and analysis of clonal heterogeneity 
4.12.6 Analysis of instability of a GS-NS0 cell line 
4.12.7 Output of transfections of GS-NS0 and GS-CHO 
4.12.8 IgG production stability of candidate GS-NS0 clones 
4.12.9 IgG production stability of GS-CHO transfectants 
4.12.10 Fed-batch bioreactor process for GS-NS0 and GS-CHO 
4.12.11 Analysis of IgG quality produced from GS-CHO and GS-NS0 bioreactor processes 
4.12.12 Comparative yield of different human IgGs produced from CHO and NS0 cells 
4.13 Summary 
5 Cell culture media development: customization of animal origin-free components and supplements 
Stephen Gorfien
5.1 Introduction 
5.2 Types of cell culture media 
5.3 Components of animal origin 
5.3.1 Segregate 
5.3.2 Mitigate 
5.3.3 Replace 
5.4 Summary and considerations for the future 
6 Post-translational modification of recombinant proteins 
Roy Jeffries
6.1 Introduction 
6.2 Common post-translational modifications 
6.3 Recombinant antibody therapeutics 
6.4 Structural and functional characteristics of human antibodies 
6.5 The human IgG subclasses: Options for antibody therapeutics 
6.6 The structure of human IgG antibodies 
6.7 IgG-Fc glycosylation 
6.8 IgG-Fab glycosylation 
6.9 Cell engineering to influence glycoform profiles 
6.10 IgG glycoforms and Fc effector functions 
6.11 Glycosylation engineering 
6.12 Pharmacokinetics and placental transport 
6.13 Antibody therapeutics of the IgA class 
6.14 Non-antibody recombinant (glyco)protein therapeutics, 'biosimilar' and 'follow-on' biologics 
6.14.1 Erythropoietin 
6.14.2 Tissue-type plasminogen activator 
6.14.3 Granulocyte-macrophage colony stimulating factor (GM-CSF) 
6.14.4 Granulocyte-colony stimulating factor 
6.14.5 Activated protein C 
6.15 Conclusions 
7 Metabolic engineering to control glycosylation 
Amy Shen, Domingos Ng, John Joly, Brad Snedecor, Yanmei Lu, Gloria Meng, Gerald Nakamura and Lynne Krummen
7.1 Introduction 
7.2 Manipulation of fucose content using RNAi technology in CHO cells 
7.2.1 Metabolic engineering of fucose content with an existing antibody production line 
7.2.2 Metabolic engineering of fucose content with simultaneous new stable cell line generation 
7.2.3 Effect of fucosylation levels on Fc_R binding 
7.2.4 Effects of fucose content on antibody-dependent cellular cytotoxicity 
7.3 Discussion 
8 An alternative approach: Humanization of N-glycosylation pathways in yeast 
Stefan Wildt and Thomas Potgieter
8.1 Introduction 
8.2 Yeast as host for recombinant protein expression 
8.3 N-linked glycosylation overview: Fungal versus mammalian 
8.4 A brief history of efforts to humanize N-linked glycosylation in fungal systems 
8.5 Sequential targeting of glycosylation enzymes is a key factor 
8.6 Replication of human-like glycosylation in the methylotrophic yeast Pichia pastoris 
8.7 A library of _-1,2 mannosidases 
8.8 Transfer of N-acetylglucosamine 
8.9 Two independent approaches towards complex N-glycans: How to eliminate more mannoses 
8.10 Some metabolic engineering: Transfer of galactose 
8.11 More metabolic engineering: Sialic acid transfer. The final step 
8.12 Glyco-engineered yeast as a host for production of therapeutic glycoproteins 
8.13 N-linked glycans and pharmacokinetics of therapeutic glycoproteins 
8.14 N-glycans and their role in tissue targeting of glycoproteins 
8.15 N-glycans can modulate the biological activity of therapeutic glycoproteins 
8.16 Control of N-glycosylation offers advantages 
8.17 Conclusions 
9 Perfusion or fed-batch? A matter of perspective 
Marco Cacciuttolo
9.1 Introduction 
9.2 Factors affecting the decision on choosing the manufacturing technology 
9.2.1 Technology expertise 
9.2.2 Facility design and scope (product dedicated versus multi-product) 
9.3 Impact of switching from perfusion to fed-batch 000
9.3.1 Personnel requirements 
9.3.2 Liquid handling 
9.3.3 Equipment 
9.3.4 Manufacturing space 
9.3.5 Decrease in cycle time 
9.3.6 Direct costs of manufacturing 
9.3.7 Productivity and morale 
9.4 Conclusions 

Library of Congress Subject Headings for this publication:

Cell culture -- Congresses.
Biochemical engineering -- Congresses.
Biotechnology -- Congresses.
Cell Culture Techniques -- methods -- Congresses.
Genetic Engineering -- methods -- Congresses.