Recombinant protein expression in mammalian cells: methods and protocols
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Bibliographic Information
Recombinant protein expression in mammalian cells: methods and protocols
（Methods in molecular biology / John M. Walker, series editor, 1850）（Springer protocols）
Humana Press, c2018
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Includes bibliographical references and index
Description and Table of Contents
Description
This detailed volume explores advances in vector design, DNA delivery, cell cultivation, host cell engineering, and bioprocess optimization within the study of recombinant protein expression in mammalian cells. The majority of the protocols employ either Chinese hamster ovary cells (CHO) or human embryonic kidney 293 cells (HEK293), the workhorses of the field, as the production host; however, the methods can be adapted to other mammalian hosts under the appropriate cell-specific conditions. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and convenient, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols aims to aid researchers in building on our knowledge of protein structure and function and to speed the discovery of new therapeutic proteins.
Table of Contents
1. Transient Gene Expression in Suspension HEK293-EBNA1 Cells
Denis L'Abbe, Louis Bisson, Christian Gervais, Eric Grazzini and Yves Durocher
2. Transient Expression of Recombinant Membrane-eGFP Fusion Proteins in HEK293 Cells
Joanna Pieprzyk, Samuel Pazicky and Christian Loew
3. PEI-Mediated Transient Gene Expression in CHO Cells
Yashas Rajendra
4. Stable Expression by Lentiviral Transduction of Cells
Natascha Goedecke, Hansjoerg Hauser and Dagmar Wirth
5. Inducible Protein Production in 293 Cells using the piggyBac Transposon System
Iacovos P. Michael and Andras Nagy
6. Recombinant CHO Cell Pool Generation using piggyBac Transposon System
Sowmya Balasubramanian
7. Genome Engineering of Hybridomas to Generate Stable Cell Lines for Antibody Expression
Cristina Parola, Derek Mason, Andreas Zingg, Daniel Neumeier and Sai T. Reddy
8. Protein Expression via Transient Transfection of Mammalian Cells in a WAVE Bioreactor
David I. Fisher
9. CHO and HEK293 Cell Cultivation and Transfection in Single-Use Orbitally Shaken Bioreactors
David L. Hacker, Laurence Durrer and Soraya Quinche
10. Bench-Scale Stirred-Tank Bioreactor for Recombinant Protein Production in Chinese Hamster Ovary Cells in Suspension
Dominique Monteil and Jeffrey Kuan
11. Continuous and Integrated Expression and Purification of Recombinant Antibodies
Sebastian Vogg, Moritz Wolf and Massimo Morbidelli
12. High Throughput Transfection of HEK293 Cells for Transient Protein Production
Tia A. Arena, Peter D. Harms and Athena W. Wong
13. Microfluidic Transfection for High-Throughput Mammalian Protein Expression
Kristina Woodruff and Sebastian J. Maerkl
14. Genome Wide High Throughput RNAi Screening for Identification of Genes Involved in Protein Production
Sarah Inwood, Michael Betenbaugh, Madhu Lal and Joseph Shiloach
15. Targeting miRNAs with CRISPR/Cas9 to Improve Recombinant Protein Production of CHO Cells
Kevin Kellner, Ankur Solanki, Thomas Amann, Nga T Lao and Niall Barron
16. Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells
Qiong Wang, Cheng-Yu Chung, Julian N. Rosenberg, Geng Yu and Michael J. Betenbaugh
17. Scalable Production and Purification of Adeno-Associated Viral Vectors (AAV)
Daniel Blessing, Nicole Deglon and Bernard L. Schneider
18. Considerations in the Use of Codon Optimization for Recombinant Protein Expression
Vincent P. Mauro and Stephen A. Chappell
19. Versatile Cell-free Protein Synthesis Systems based on Chinese Hamster Ovary Cells
Lena Thoring and Stefan Kubick
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